Anti-Calnexin antibody [CANX/1543]

35 product images

• 

  Western blot - Anti-Calnexin antibody [CANX/1543] (ab238078)

  Lanes 1 - 4: Merged signal (red and green). Green - ab238078 observed at 80 kDa. Red - loading control Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) observed at 37 kDa.
  

  ab238078 was shown to react with Calnexin in wild-type HEK-293T cells in Western blot with loss of signal observed in CANX knockout cell line Human CANX (Calnexin) knockout HEK-293T cell line ab255368 (CANX knockout cell lysate Human CANX (Calnexin) knockout HEK-293T cell lysate ab263805). Wild-type HEK-293T and CANX knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween^®) before incubation with ab238078 and Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) overnight at 4 °C at 1 μg/ml and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye^® 800CW) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) and Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preabsorbed (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

  All lanes: Western blot - Anti-Calnexin antibody [CANX/1543] (ab238078) at 1 µg/mL

  Lane 1: Wild-type HEK-293T cell lysate at 20 µg

  Lane 2: CANX knockout HEK-293T cell lysate at 20 µg

  Lane 3: U-2 OS cell lysate at 20 µg

  Lane 4: MCF7 cell lysate at 20 µg

  Performed under reducing conditions.

  Predicted band size: 68 kDa

  Observed band size: 80 kDa

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calnexin antibody [CANX/1543] (ab238078)

  Formalin-fixed, paraffin-embedded human renal cell carcinoma tissue stained for Calnexin using ab238078 at 2 μg/ml in immunohistochemical analysis.

• 

  Protein Array - Anti-Calnexin antibody [CANX/1543] (ab238078)

  ab238078 was tested in protein array against over 19000 different full-length human proteins.
  Z- and S- Score: The Z-score represents the strength of a signal that a monoclonal antibody (MAb) (in combination with a fluorescently-tagged anti-IgG secondary antibody) produces when binding to a particular protein on the HuProtTM array. Z-scores are described in units of standard deviations (SD's) above the mean value of all signals generated on that array. If targets on HuProtTM are arranged in descending order of the Z-score, the S-score is the difference (also in units of SD's) between the Z-score. S-score therefore represents the relative target specificity of a MAb to its intended target.
  A MAb is specific to its intended target if the MAb has an S-score of at least 2.5. For example, if a MAb binds to protein X with a Z-score of 43 and to protein Y with a Z-score of 14, then the S-score for the binding of that MAb to protein X is equal to 29.

• 

  Western blot - Anti-Calnexin antibody [CANX/1543] (ab238078)

  All lanes: Western blot - Anti-Calnexin antibody [CANX/1543] (ab238078) at 2 µg/mL

  Lane 1: PANC-1 (human pancreatic epithelial cancinoma cell line) cell lysate

  Lane 2: MCF7 (human breast adenocarcinoma cell line) cell lysate

  Predicted band size: 68 kDa

• 

  Western blot - Anti-Calnexin antibody [CANX/1543] (ab238078)

  All lanes: Western blot - Anti-Calnexin antibody [CANX/1543] (ab238078) at 2 µg/mL

  All lanes: Human kidney lysate

  Predicted band size: 68 kDa

• 

  Western blot - Anti-Calnexin antibody [CANX/1543] (ab238078)

  Western blot: Anti-LATS2 antibody [EPR23126-488] (Anti-LATS2 antibody [EPR23126-488] ab243657) staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, Anti-LATS2 antibody [EPR23126-488] ab243657 was shown to bind specifically to LATS2. A band was observed at 117-171 kDa in wild-type A549 cell lysates with no signal observed at this size in LATS2 knockout cell line. To generate this image, wild-type and LATS2 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

  All lanes: Western blot - Anti-LATS2 antibody [EPR23126-488] (Anti-LATS2 antibody [EPR23126-488] ab243657) at 1/1000 dilution

  Lane 1: Wild-type A549 cell lysate at 20 µg

  Lane 2: LATS2 knockout A549 cell lysate at 20 µg

  Lane 3: MCF7 cell lysate at 20 µg

  Lane 4: Raji cell lysate at 20 µg

  Secondary

  Lanes 1 - 4: Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution

  Lanes 1 - 4: Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

  Performed under reducing conditions.

  Observed band size: 117-171 kDa

• 

  Western blot - Anti-Calnexin antibody [CANX/1543] (ab238078)

  Western blot: Anti-PTPN2 antibody [EPR28199-34] (Anti-TCPTP antibody [EPR28199-34] ab314496) staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, Anti-TCPTP antibody [EPR28199-34] ab314496 was shown to bind specifically to PTPN2. A band was observed at 48 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in PTPN2 knockout cell line. To generate this image, wild-type and PTPN2 knockout HCT 116 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

  All lanes: Western blot - Anti-TCPTP antibody [EPR28199-34] (Anti-TCPTP antibody [EPR28199-34] ab314496) at 1/1000 dilution

  Lane 1: Wild-type HCT 116 cell lysate at 20 µg

  Lane 2: PTPN2 knockout HCT 116 cell lysate at 20 µg

  Lane 3: Wild-type Jurkat cell lysate at 20 µg

  Lane 4: PTPN2 knockout Jurkat cell lysate at 20 µg

  Secondary

  Lanes 1 - 4: Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution

  Lanes 1 - 4: Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

  Performed under reducing conditions.

  Observed band size: 48 kDa

• 

  Western blot - Anti-Calnexin antibody [CANX/1543] (ab238078)

  Western blot: Anti-RB1 antibody (Anti-Rb antibody ab226979) staining at 1/1500 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, Anti-Rb antibody ab226979 was shown to bind specifically to RB1. A band was observed at 120 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in RB1 knockout cell line. To generate this image, wild-type and RB1 knockout HCT 116 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

  All lanes: Western blot - Anti-Rb antibody (Anti-Rb antibody ab226979) at 1/1500 dilution

  Lane 1: Wild-type HCT 116 cell lysate at 20 µg

  Lane 2: RB1 knockout HCT 116 cell lysate at 20 µg

  Lane 3: Wild-type A549 Human wild-type A549 cell line ab288558 cell lysate at 20 µg

  Lane 4: RB1 knockout A549 Human RB1 knockout A549 cell line ab286470 cell lysate at 20 µg

  Secondary

  Lanes 1 - 4: Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution

  Lanes 1 - 4: Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

  Performed under reducing conditions.

  Observed band size: 120 kDa

• 

  Western blot - Anti-Calnexin antibody [CANX/1543] (ab238078)

  Western blot: Anti-PIKFYVE antibody (Anti-PIP5K3/PIKFYVE antibody [EPR27944-21] ab315090) staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, Anti-PIP5K3/PIKFYVE antibody [EPR27944-21] ab315090 was shown to bind specifically to PIKFYVE. A band was observed at 237 kDa in wild-type A549 cell lysates with no signal observed at this size in PIKFYVE knockout cell line. To generate this image, wild-type and PIKFYVE knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

  All lanes: Western blot - Anti-PIP5K3/PIKFYVE antibody [EPR27944-21] (Anti-PIP5K3/PIKFYVE antibody [EPR27944-21] ab315090) at 1/1000 dilution

  Lane 1: Wild-type A549 cell lysate at 20 µg

  Lane 2: PIKFYVE knockout A549 cell lysate at 20 µg

  Lane 3: HeLa UV treated (100 mJ/cm^2, 1 hr recovery, new, Branford) cell lysate at 20 µg

  Lane 4: HeLa UV treated (40 mJ/cm^2 30 min recovery), LAMBDA PHOS 4X, 2h cell lysate at 20 µg

  Secondary

  Lanes 1 - 4: Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution

  Lanes 1 - 4: Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

  Performed under reducing conditions.

  Observed band size: 237 kDa

• 

  Western blot - Anti-Calnexin antibody [CANX/1543] (ab238078)

  All lanes: Western blot - Anti-KDM6A / UTX antibody [EPR26387-23] (Anti-KDM6A / UTX antibody [EPR26387-23] ab300513) at 1/1000 dilution

  Lane 1: Wild-type A549 cell lysate at 7 µg

  Lane 2: KDM6A/UTX knockout A549 cell lysate at 7 µg

  Secondary

  Lanes 1 - 2: Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution

  Lanes 1 - 2: Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

  Performed under reducing conditions.

  Observed band size: 154 kDa

• 

  Western blot - Anti-Calnexin antibody [CANX/1543] (ab238078)

  Western blot: Anti-DROSHA antibody [EPR25336-93] (Anti-Drosha antibody [EPR25336-93] ab303544) staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, Anti-Drosha antibody [EPR25336-93] ab303544 was shown to bind specifically to DROSHA. A band was observed at 117-171 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in DROSHA knockout cell line. To generate this image, wild-type and DROSHA knockout HCT 116 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

  All lanes: Western blot - Anti-Drosha antibody [EPR25336-93] (Anti-Drosha antibody [EPR25336-93] ab303544) at 1/1000 dilution

  Lane 1: Wild-type HCT 116 cell lysate at 20 µg

  Lane 2: DROSHA knockout HCT 116 cell lysate at 20 µg

  Secondary

  Lanes 1 - 2: Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution

  Lanes 1 - 2: Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

  Performed under reducing conditions.

  Observed band size: 117-171 kDa

• 

  Western blot - Anti-Calnexin antibody [CANX/1543] (ab238078)

  Western blot: Anti-CEP290 antibody (Anti-CEP290 antibody ab85728) staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, Anti-CEP290 antibody ab85728 was shown to bind specifically to CEP290. A band was observed at 290 kDa in wild-type A549 cell lysates with no signal observed at this size in CEP290 knockout cell line. To generate this image, wild-type and CEP290 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

  All lanes: Western blot - Anti-CEP290 antibody (Anti-CEP290 antibody ab85728) at 1/1000 dilution

  Lane 1: Wild-type A549 cell lysate at 40 µg

  Lane 2: CEP290 knockout A549 cell lysate at 20 µg

  Lane 3: Wild-type A549 Human wild-type A549 cell line ab288558 cell lysate at 10 µg

  Lane 4: CEP290 knockout A549 STN-436528 cell lysate at 40 µg

  Secondary

  Lanes 1 - 4: Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution

  Lanes 1 - 4: Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

  Performed under reducing conditions.

  Observed band size: 290 kDa

• 

  Western blot - Anti-Calnexin antibody [CANX/1543] (ab238078)

  Western blot: Anti-CEP290 antibody (Anti-CEP290 antibody ab84870) staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, Anti-CEP290 antibody ab84870 was shown to bind specifically to CEP290. A band was observed at 290 kDa in wild-type A549 cell lysates with no signal observed at this size in CEP290 knockout cell line. To generate this image, wild-type and CEP290 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

  All lanes: Western blot - Anti-CEP290 antibody (Anti-CEP290 antibody ab84870) at 1/1000 dilution

  Lane 1: Wild-type A549 cell lysate at 20 µg

  Lane 2: CEP290 knockout A549 cell lysate at 20 µg

  Lane 3: Wild-type A549 Human wild-type A549 cell line ab288558 cell lysate at 10 µg

  Lane 4: CEP290 knockout A549 STN-436528 cell lysate at 40 µg

  Secondary

  Lanes 1 - 4: Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution

  Lanes 1 - 4: Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

  Performed under reducing conditions.

  Observed band size: 290 kDa

• 

  Western blot - Anti-Calnexin antibody [CANX/1543] (ab238078)

  Western blot: Anti-FAT1 antibody (Anti-FAT/FAT1 antibody ab241372) staining at 1/2000 dilution, shown in black; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, Anti-FAT/FAT1 antibody ab241372 was shown to bind specifically to FAT1. A band was observed at 506 kDa in wild-type A549 cell lysates with no signal observed at this size in FAT1 knockout cell line. To generate this image, wild-type and FAT1 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times before development with a high-sensitivity ECL substrate kit and imaged with 3 minutes exposure time. Secondary antibodies used were HRP conjugated Goat anti-Rabbit (H+L) and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

  All lanes: Western blot - Anti-FAT/FAT1 antibody (Anti-FAT/FAT1 antibody ab241372) at 1/2000 dilution

  Lane 1: Wild-type A549 cell lysate at 20 µg

  Lane 2: FAT1 knockout A549 cell lysate at 20 µg

  Lane 3: Jurkat cell lysate at 20 µg

  Lane 4: HUVEC cell lysate at 20 µg

  Secondary

  Lanes 1 - 4: HRP conjugated Goat anti-Rabbit (H+L) at 1/20000 dilution

  Lanes 1 - 4: Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

  Performed under reducing conditions.

  Observed band size: 506 kDa

• 

  Western blot - Anti-Calnexin antibody [CANX/1543] (ab238078)

  Western blot: Anti-ESRRA antibody (Anti-Estrogen Related Receptor alpha antibody ab137489) staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, Anti-Estrogen Related Receptor alpha antibody ab137489 was shown to bind specifically to ESRRA. A band was observed at 46 kDa in wild-type MCF7 cell lysates with no signal observed at this size in ESRRA knockout cell line. To generate this image, wild-type and ESRRA knockout MCF7 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

  All lanes: Western blot - Anti-Estrogen Related Receptor alpha antibody (Anti-Estrogen Related Receptor alpha antibody ab137489) at 1/1000 dilution

  Lane 1: Wild-type MCF7 cell lysate at 20 µg

  Lane 2: ESRRA knockout MCF7 cell lysate at 20 µg

  Lane 3: Wild-type HAP1 cell lysate at 20 µg

  Lane 4: ESRRA knockout HAP1 cell lysate at 20 µg

  Secondary

  Lanes 1 - 4: Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution

  Lanes 1 - 4: Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

  Performed under reducing conditions.

  Observed band size: 46 kDa

• 

  Western blot - Anti-Calnexin antibody [CANX/1543] (ab238078)

  Western blot: Anti-ESRRA antibody [EPR46Y] (Anti-Estrogen Related Receptor alpha antibody [EPR46Y] ab76228) staining at 1/2000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, Anti-Estrogen Related Receptor alpha antibody [EPR46Y] ab76228 was shown to bind specifically to ESRRA. A band was observed at 46 kDa in wild-type MCF7 cell lysates with no signal observed at this size in ESRRA knockout cell line. To generate this image, wild-type and ESRRA knockout MCF7 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

  All lanes: Western blot - Anti-Estrogen Related Receptor alpha antibody [EPR46Y] (Anti-Estrogen Related Receptor alpha antibody [EPR46Y] ab76228) at 1/2000 dilution

  Lane 1: Wild-type MCF7 cell lysate at 20 µg

  Lane 2: ESRRA knockout MCF7 cell lysate at 20 µg

  Lane 3: Wild-type HAP1 cell lysate at 20 µg

  Lane 4: ESRRA knockout HAP1 cell lysate at 20 µg

  Secondary

  Lanes 1 - 4: Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution

  Lanes 1 - 4: Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

  Performed under reducing conditions.

  Observed band size: 46 kDa

• 

  Western blot - Anti-Calnexin antibody [CANX/1543] (ab238078)

  Western blot: Anti-SMAD3 (phospho S423 + S425) antibody [EP619Y] (Anti-SMAD5 antibody [EP619Y] ab40771) staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, Anti-SMAD5 antibody [EP619Y] ab40771 was shown to bind specifically to SMAD3 (phospho S423 + S425). A band was observed at 52 kDa in wild-type HeLa cell lysates with no signal observed at this size in SMAD3 (phospho S423 + S425) knockout cell line. To generate this image, wild-type and SMAD3 (phospho S423 + S425) knockout HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

  All lanes: Western blot - Anti-SMAD5 antibody [EP619Y] (Anti-SMAD5 antibody [EP619Y] ab40771) at 1/1000 dilution

  Lane 1: Wild-type HeLa Vehicle Control TGF-beta (0 ng/mL, 30 min) cell lysate at 20 µg

  Lane 2: Wild-type HeLa Treated TGF-beta (10 ng/mL, 30 min) cell lysate at 20 µg

  Lane 3: SMAD1 knockout HeLa Vehicle Control TGF-beta (0 ng/mL, 30 min), Human SMAD1 knockout HeLa cell line ab265400 cell lysate at 20 µg

  Lane 4: SMAD1 knockout HeLa Treated TGF-beta (10 ng/mL, 30 min), Human SMAD1 knockout HeLa cell line ab265400 cell lysate at 20 µg

  Lane 5: SMAD2 knockout HeLa Vehicle Control TGF-beta (0 ng/mL, 30 min), Human SMAD2 knockout HeLa cell line ab255430 cell lysate at 20 µg

  Lane 6: SMAD2 knockout HeLa Treated TGF-beta (10 ng/mL, 30 min), Human SMAD2 knockout HeLa cell line ab255430 cell lysate at 20 µg

  Lane 7: Wild-type HeLa Vehicle Control TGF-beta (0 ng/mL, 30 min), Human wild-type HeLa cell line ab255448 cell lysate at 20 µg

  Lane 8: Wild-type HeLa Treated TGF-beta (10 ng/mL, 30 min), Human wild-type HeLa cell line ab255448 cell lysate at 20 µg

  Lane 9: SMAD3 knockout HeLa Vehicle Control TGF-beta (0 ng/mL, 30 min), Human SMAD3 knockout HeLa cell line ab255431 cell lysate at 20 µg

  Lane 10: SMAD3 knockout HeLa Treated TGF-beta (10 ng/mL, 30 min), Human SMAD3 knockout HeLa cell line ab255431 cell lysate at 20 µg

  Lane 11: Wild-type HEK293 Vehicle Control TGF-beta (0 ng/mL, 30 min), Human wild-type HEK-293 cell line ab259776 cell lysate at 20 µg

  Lane 12: Wild-type HEK293 Treated TGF-beta (10 ng/mL, 30 min), Human wild-type HEK-293 cell line ab259776 cell lysate at 20 µg

  Lane 13: SMAD5 knockout HEK293 Vehicle Control TGF-beta (0 ng/mL, 30 min), Human SMAD5 knockout HEK-293 cell line ab269470 cell lysate at 20 µg

  Lane 14: SMAD5 knockout HEK293 Treated TGF-beta (10 ng/mL, 30 min), Human SMAD5 knockout HEK-293 cell line ab269470 cell lysate at 20 µg

  Secondary

  Lanes 1, 10, 11, 12, 13, 14, 2, 3, 4, 5, 6, 7, 8 and 9: Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution

  Lanes 1, 10, 11, 12, 13, 14, 2, 3, 4, 5, 6, 7, 8 and 9: Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

  Performed under reducing conditions.

  Observed band size: 52 kDa

• 

  Western blot - Anti-Calnexin antibody [CANX/1543] (ab238078)

  Western blot: Anti-MAP2K2 antibody [Y78] (Anti-MEK2 antibody [Y78] ab32517) staining at 1/10000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, Anti-MEK2 antibody [Y78] ab32517 was shown to bind specifically to MAP2K2. A band was observed at 44 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in MAP2K2 knockout cell line. To generate this image, wild-type and MAP2K2 knockout HCT 116 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

  All lanes: Western blot - Anti-MEK2 antibody [Y78] (Anti-MEK2 antibody [Y78] ab32517) at 1/10000 dilution

  Lane 1: Wild-type HCT 116 cell lysate at 20 µg

  Lane 2: MAP2K2 knockout HCT 116 cell lysate at 20 µg

  Lane 3: Wild-type HEK-293T ab255553 cell lysate at 20 µg

  Lane 4: MAP2K2 knockout HEK-293T ab261003 cell lysate at 20 µg

  Secondary

  Lanes 1 - 4: Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution

  Lanes 1 - 4: Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

  Performed under reducing conditions.

  Observed band size: 44 kDa

• 

  Western blot - Anti-Calnexin antibody [CANX/1543] (ab238078)

  Western blot: Anti-DROSHA antibody [RM1063] (Anti-Drosha antibody [RM1063] ab315100) staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, Anti-Drosha antibody [RM1063] ab315100 was shown to bind specifically to DROSHA. A band was observed at 117-171 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in DROSHA knockout cell line. To generate this image, wild-type and DROSHA knockout HCT 116 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

  All lanes: Western blot - Anti-Drosha antibody [RM1063] (Anti-Drosha antibody [RM1063] ab315100) at 1/1000 dilution

  Lane 1: Wild-type HCT 116 cell lysate at 20 µg

  Lane 2: DROSHA knockout HCT 116 cell lysate at 20 µg

  Secondary

  Lanes 1 - 2: Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution

  Lanes 1 - 2: Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

  Performed under reducing conditions.

  Observed band size: 117-171 kDa

• 

  Western blot - Anti-Calnexin antibody [CANX/1543] (ab238078)

  Western blot: Anti-GJA1 antibody [EPR21153] (Anti-Connexin 43 / GJA1 antibody [EPR21153] ab217676) staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, Anti-Connexin 43 / GJA1 antibody [EPR21153] ab217676 was shown to bind specifically to GJA1. A band was observed at 45 kDa in wild-type U-87 MG cell lysates with no signal observed at this size in GJA1 knockout cell line. To generate this image, wild-type and GJA1 knockout U-87 MG cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

  All lanes: Western blot - Anti-Connexin 43 / GJA1 antibody [EPR21153] (Anti-Connexin 43 / GJA1 antibody [EPR21153] ab217676) at 1/1000 dilution

  Lane 1: Wild-type U-87 MG cell lysate at 20 µg

  Lane 3: Wild-type HEK-293 cell lysate at 20 µg

  Lane 4: GJA1 knockout HEK-293 Human GJA1 knockout HEK-293 cell lysate ab261658 cell lysate at 20 µg

  Secondary

  Lanes 1 - 4: Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution

  Lanes 1 - 4: Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

  Performed under reducing conditions.

  Observed band size: 45 kDa

• 

  Western blot - Anti-Calnexin antibody [CANX/1543] (ab238078)

  Western blot: Anti-TTF1/Nkx2-1 antibody [EP1584Y] (Biotin Anti-TTF1/Nkx2-1 antibody [EP1584Y] ab314937) staining at 1/2000 dilution, shown in black; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta.

  In Western blot, Biotin Anti-TTF1/Nkx2-1 antibody [EP1584Y] ab314937 was shown to bind specifically to NKX2-1/TTF-1. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5% milk in TBS-0.1% Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times before development with a high-sensitivity ECL substrate kit.

  Secondary antibodies used were HRP conjugated Streptavidin at 0.1 µg/ml and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

  All lanes: Western blot - Biotin Anti-TTF1/Nkx2-1 antibody [EP1584Y] (Biotin Anti-TTF1/Nkx2-1 antibody [EP1584Y] ab314937) at 1/2000 dilution

  Lane 1: Hela cell lysate at 20 µg

  Lane 2: HEK-293 cell lysate at 20 µg

  Secondary

  Lane 1: HRP conjugated Streptavidin at 0.1 µg/mL

  Lane 2: Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

  Developed using the ECL technique.

  Performed under reducing conditions.

  Observed band size: 39 kDa, 75 kDa

  Exposure time: 20min

• 

  Western blot - Anti-Calnexin antibody [CANX/1543] (ab238078)

  Anti-NEFM antibody [EP2460] (Anti-160 kD Neurofilament Medium antibody [EP2460] ab92539) staining at 1/100000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, Anti-160 kD Neurofilament Medium antibody [EP2460] ab92539 was shown to bind specifically to NEFM. A band was observed at 140-160 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in NEFM knockout cell line. To generate this image, wild-type and NEFM knockout HEK-293T cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween$^®$ 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

  All lanes: Western blot - Anti-160 kD Neurofilament Medium antibody [EP2460] (Anti-160 kD Neurofilament Medium antibody [EP2460] ab92539) at 1/100000 dilution

  Lane 1: Wild-type HEK-293T cell lysate at 20 µg

  Lane 2: NEFM knockout HEK-293T cell lysate at 20 µg

  Lane 3: SK-N-BE cell lysate at 20 µg

  Lane 4: A549 cell lysate at 20 µg

  Lane 5: SH-SY5Y cell lysate at 20 µg

  Secondary

  Lanes 1 - 5: Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution

  Lanes 1 - 5: Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

  Predicted band size: 102 kDa

  Observed band size: 140-160 kDa

• 

  Western blot - Anti-Calnexin antibody [CANX/1543] (ab238078)

  Western blot: Anti-CEACAM5 antibody (Anti-Carcino Embryonic Antigen CEA antibody ab131070) staining at 0.1 ug/ml, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-Carcino Embryonic Antigen CEA antibody ab131070 was shown to bind specifically to CEACAM5. A band was observed at 170 kDa in wild-type A549 cell lysates with no signal observed at this size in CEACAM5 knockout cell line. To generate this image, wild-type and CEACAM5 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

  All lanes: Western blot - Anti-Carcino Embryonic Antigen CEA antibody (Anti-Carcino Embryonic Antigen CEA antibody ab131070) at 0.1 µg/mL

  Lane 1: Wild-type A549 cell lysate at 20 µg

  Lane 2: CEACAM5 knockout A549 cell lysate at 20 µg

  Lane 3: MCF7 cell lysate at 20 µg

  Lane 4: PANC-1 cell lysate at 20 µg

  Performed under reducing conditions.

  Predicted band size: 76 kDa

• 

  Western blot - Anti-Calnexin antibody [CANX/1543] (ab238078)

  False colour image of Western blot: Anti-DNA PKcs antibody [Y393] staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-DNA PKcs antibody [Y393] ab32566 was shown to bind specifically to DNA PKcs. A band was observed at 450 kDa in wild-type A549 cell lysates with no signal observed at this size in PRKDC knockout cell line. To generate this image, wild-type and PRKDC knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

  All lanes: Western blot - Anti-DNA PKcs antibody [Y393] (Anti-DNA PKcs antibody [Y393] ab32566) at 1/1000 dilution

  Lane 1: Wild-type A549 cell lysate at 20 µg

  Lane 2: PRKDC knockout A549 cell lysate at 20 µg

  Lane 3: K562 cell lysate at 20 µg

  Lane 4: HDLM-2 cell lysate at 20 µg

  Secondary

  All lanes: Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

  Performed under reducing conditions.

  Predicted band size: 469 kDa

  Observed band size: 450 kDa

• 

  Western blot - Anti-Calnexin antibody [CANX/1543] (ab238078)

  Western blot: HRP Anti-ALDH5A1/SSADH antibody [EPR7794] (HRP Anti-ALDH5A1/SSADH antibody [EPR7794] ab314408) staining at 1/1000 dilution, shown in black; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in red. In Western blot, HRP Anti-ALDH5A1/SSADH antibody [EPR7794] ab314408 was shown to bind specifically to ALDH5A1. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibody Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution for 1 h at room temperature, washed again four times before development with a high-sensitivity ECL substrate kit and imaged with 8 minutes exposure time.

  All lanes: Western blot - HRP Anti-ALDH5A1/SSADH antibody [EPR7794] (HRP Anti-ALDH5A1/SSADH antibody [EPR7794] ab314408) at 1/1000 dilution

  Lane 1: A431 cell lysate at 20 µg

  Lane 2: Human liver lysate at 20 µg

  Secondary

  All lanes: Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

  Performed under reducing conditions.

  Observed band size: 54 kDa

• 

  Western blot - Anti-Calnexin antibody [CANX/1543] (ab238078)

  Western blot: Anti-EIF2AK3 antibody [EPR19876-294] (Anti-PERK antibody [EPR19876-294] ab229912) staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, Anti-PERK antibody [EPR19876-294] ab229912 was shown to bind specifically to EIF2AK3. A band was observed at 145 kDa in wild-type A549 cell lysates with no signal observed at this size in EIF2AK3 knockout cell line. To generate this image, wild-type and EIF2AK3 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

  All lanes: Western blot - Anti-PERK antibody [EPR19876-294] (Anti-PERK antibody [EPR19876-294] ab229912) at 1/1000 dilution

  Lane 1: Wild-type A549 cell lysate at 20 µg

  Lane 2: EIF2AK3 knockout A549 cell lysate at 20 µg

  Lane 3: MCF7 cell lysate at 20 µg

  Lane 4: Human eye cell lysate at 20 µg

  Secondary

  All lanes: Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

  Performed under reducing conditions.

  Observed band size: 145 kDa

• 

  Western blot - Anti-Calnexin antibody [CANX/1543] (ab238078)

  Western blot: Anti-ALDOA antibody [EPR23181-39] (Anti-Aldolase antibody [EPR23181-39] ab252953) staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, Anti-Aldolase antibody [EPR23181-39] ab252953 was shown to bind specifically to ALDOA. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

  All lanes: Western blot - Anti-Aldolase antibody [EPR23181-39] (Anti-Aldolase antibody [EPR23181-39] ab252953) at 1/1000 dilution

  Lane 1: HEK-293T overexpressing ALDOA cell lysate at 20 µg

  Lane 2: HEK-293T control cell lysate at 20 µg

  Lane 3: MCF7 cell lysate at 20 µg

  Lane 4: HeLa cell lysate at 20 µg

  Lane 5: Human heart cell lysate at 20 µg

  Performed under reducing conditions.

  Observed band size: 45 kDa

• 

  Western blot - Anti-Calnexin antibody [CANX/1543] (ab238078)

  Western blot: Anti-EP300 antibody [EPR23495-268] (Anti-KAT3B / p300 antibody [EPR23495-268] - ChIP Grade ab275378) staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, Anti-KAT3B / p300 antibody [EPR23495-268] - ChIP Grade ab275378 was shown to bind specifically to EP300. A band was observed at 200/90-130 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in EP300 knockout cell line. To generate this image, wild-type and EP300 knockout HCT 116 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

  All lanes: Western blot - Anti-KAT3B / p300 antibody [EPR23495-268] - ChIP Grade (Anti-KAT3B / p300 antibody [EPR23495-268] - ChIP Grade ab275378) at 1/1000 dilution

  Lane 1: Wild-type HCT 116 cell lysate at 20 µg

  Lane 2: EP300 knockout HCT 116 cell lysate at 20 µg

  Lane 3: HeLa cell lysate at 20 µg

  Lane 4: Human brain olfactory lobe cell lysate at 20 µg

  Secondary

  All lanes: Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

  Performed under reducing conditions.

  Observed band size: 200 kDa, 90-130 kDa

• 

  Western blot - Anti-Calnexin antibody [CANX/1543] (ab238078)

  Western blot: Anti-CBL antibody [YE323] (Anti-CBL antibody [YE323] - C-terminal ab32027) staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, Anti-CBL antibody [YE323] - C-terminal ab32027 was shown to bind specifically to CBL. A band was observed at 105 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in CBL knockout cell line. To generate this image, wild-type and CBL knockout HCT 116 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

  All lanes: Western blot - Anti-CBL antibody [YE323] - C-terminal (Anti-CBL antibody [YE323] - C-terminal ab32027) at 1/1000 dilution

  Lane 1: Wild-type HCT 116 cell lysate at 20 µg

  Lane 2: CBL knockout HCT 116 cell lysate at 20 µg

  Secondary

  All lanes: Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

  Performed under reducing conditions.

  Observed band size: 105 kDa

• 

  Western blot - Anti-Calnexin antibody [CANX/1543] (ab238078)

  Western blot: Anti-INPPL1 antibody [EPR10954] (Anti-INPPL1/SHIP-2 antibody [EPR10954] ab157460) staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, Anti-INPPL1/SHIP-2 antibody [EPR10954] ab157460 was shown to bind specifically to INPPL1. A band was observed at 110-150 kDa in wild-type A549 cell lysates with no signal observed at this size in INPPL1 knockout cell line. To generate this image, wild-type and INPPL1 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

  All lanes: Western blot - Anti-INPPL1/SHIP-2 antibody [EPR10954] (Anti-INPPL1/SHIP-2 antibody [EPR10954] ab157460) at 1/1000 dilution

  Lane 1: Wild-type A549 cell lysate at 20 µg

  Lane 2: INPPL1 knockout A549 cell lysate at 20 µg

  Secondary

  All lanes: Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

  Performed under reducing conditions.

  Observed band size: 110-150 kDa

• 

  Western blot - Anti-Calnexin antibody [CANX/1543] (ab238078)

  Western blot: Anti-EP300 antibody (Anti-KAT3B / p300 antibody ab10485) staining at 1/15000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, Anti-KAT3B / p300 antibody ab10485 was shown to bind specifically to EP300. A band was observed at 200/90-130 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in EP300 knockout cell line. To generate this image, wild-type and EP300 knockout HCT 116 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

  All lanes: Western blot - Anti-KAT3B / p300 antibody (Anti-KAT3B / p300 antibody ab10485) at 1/15000 dilution

  Lane 1: Wild-type HCT 116 cell lysate at 20 µg

  Lane 2: EP300 knockout HCT 116 cell lysate at 20 µg

  Lane 3: HeLa cell lysate at 20 µg

  Lane 4: Human brain olfactory lobe cell lysate at 20 µg

  Secondary

  All lanes: Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

  Performed under reducing conditions.

  Observed band size: 200 kDa, 90-130 kDa

• 

  Western blot - Anti-Calnexin antibody [CANX/1543] (ab238078)

  Western blot: Anti-TET2 antibody (Anti-Tet2 antibody ab124297) staining at 1/250 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, Anti-Tet2 antibody ab124297 was shown to bind specifically to TET2. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

  All lanes: Western blot - Anti-Tet2 antibody (Anti-Tet2 antibody ab124297) at 1/250 dilution

  Lane 1: mES cell lysate at 20 µg

  Lane 2: F9 cell lysate at 20 µg

  Lane 3: NIH/3T3 cell lysate at 20 µg

  Performed under reducing conditions.

  Observed band size: 225 kDa

• 

  Western blot - Anti-Calnexin antibody [CANX/1543] (ab238078)

  Western blot: Anti-EIF2AK3 antibody (Anti-PERK antibody ab65142) staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, Anti-PERK antibody ab65142 was shown to bind specifically to EIF2AK3. A band was observed at 145 kDa in wild-type A549 cell lysates with no signal observed at this size in EIF2AK3 knockout cell line. To generate this image, wild-type and EIF2AK3 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

  All lanes: Western blot - Anti-PERK antibody (Anti-PERK antibody ab65142) at 1/1000 dilution

  Lane 1: Wild-type A549 cell lysate at 20 µg

  Lane 2: EIF2AK3 knockout A549 cell lysate at 20 µg

  Lane 3: MCF7 cell lysate at 20 µg

  Lane 4: Human eye cell lysate at 20 µg

  Secondary

  All lanes: Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

  Performed under reducing conditions.

  Observed band size: 145 kDa

• 

  Western blot - Anti-Calnexin antibody [CANX/1543] (ab238078)

  Western blot: Anti-EHMT1 antibody [EPR18667] (Anti-EHMT2/G9A + EHMT1/GLP antibody [EPR18667] ab194299) staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, Anti-EHMT2/G9A + EHMT1/GLP antibody [EPR18667] ab194299 was shown to bind specifically to EHMT1. A band was observed at 165 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in EHMT1 knockout cell line. To generate this image, wild-type and EHMT1 knockout HCT 116 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

  All lanes: Western blot - Anti-EHMT2/G9A + EHMT1/GLP antibody [EPR18667] (Anti-EHMT2/G9A + EHMT1/GLP antibody [EPR18667] ab194299) at 1/1000 dilution

  Lane 1: Wild-type HCT 116 cell lysate at 20 µg

  Lane 2: EHMT1 knockout HCT 116 cell lysate at 20 µg

  Secondary

  All lanes: Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

  Performed under reducing conditions.

  Observed band size: 165 kDa

• 

  Western blot - Anti-Calnexin antibody [CANX/1543] (ab238078)

  All lanes: Anti-USP18 antibody at 1/1000 dilution

  Lane 1: Wild-type A549 Vehicle Control IFNa (0 IU/mL, 12 h) cell lysate at 20 µg

  Lane 2: Wild-type A549 Treated IFNa (100 IU/mL, 12 h) cell lysate at 20 µg

  Lane 3: USP18 knockout A549 Vehicle Control IFNa (0 IU/mL, 12 h) ab262530 cell lysate at 20 µg

  Lane 4: USP18 knockout A549 Treated IFNa (100 IU/mL, 12 h) ab262530 cell lysate at 20 µg

  Secondary

  Lanes 1 - 4: Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution

  Lanes 1 - 4: Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

  Predicted band size: 43 kDa

  Observed band size: 34-37 kDa