Anti-CD167a/DDR1 antibody [EPR24783-7]

• IP

Lab

Immunoprecipitation - Anti-CD167a/DDR1 antibody [EPR24783-7] (AB288675)

Epithelial discoidin domain-containing receptor 1 was immunoprecipitated from 0.5 mg T-47D lysates with 5 μg ab288675. Western blot was performed on the immunoprecipitate using ab288675 at 1/1000.

Lane 1 : T-47D lysate at 10 µg
Lane 2 : T-47D Pulldown with 10 µg/mL ab288675
Lane 3 : T-47D Pulldown with 10 µg/mL ab172730 isotype control

Observed band size : 120 kDa

For Western blot, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % NFDM 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 in 3 % NFDM block, washed again four times then imaged.

All lanes:

Immunoprecipitation - Anti-CD167a/DDR1 antibody [EPR24783-7] (ab288675) at 10 µg/mL

All lanes:

T-47D lysate with 3 % NFDM in TBS-0.1% Tween® 20 (TBS-T)

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/1000 dilution

false

Exposure time: 20min

• IP

Lab

Immunoprecipitation - Anti-CD167a/DDR1 antibody [EPR24783-7] (AB288675)

Epithelial discoidin domain-containing receptor 1 was immunoprecipitated from 0.5 mg NIH/3T3 lysates with 5 μg ab288675. Western blot was performed on the immunoprecipitate using ab288675 at 1/1000.

Lane 1 : NIH/3T3 lysate at 10 µg
Lane 2 : NIH/3T3 Pulldown with 10 µg/mL ab288675
Lane 3 : NIH/3T3 Pulldown with 10 µg/mL ab172730 isotype control

Observed band size : 120 kDa

For Western blot, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % NFDM 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 in 3 % NFDM block, washed again four times then imaged.

All lanes:

Immunoprecipitation - Anti-CD167a/DDR1 antibody [EPR24783-7] (ab288675) at 10 µg/mL

All lanes:

NIH/3T3 lysate with 3 % NFDM in TBS-0.1% Tween® 20 (TBS-T)

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/1000 dilution

false

Exposure time: 4min

• WB

Lab

Western blot - Anti-CD167a/DDR1 antibody [EPR24783-7] (AB288675)

Western blot : Anti-DDR1 antibody [EPR24783-7] (ab288675) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab288675 was shown to bind specifically to DDR1. A band was observed at 100-125 kDa in wild-type HeLa cell lysates with no signal observed at this size in DDR1 knockout cell line ab280048 (knockout cell lysate ab280107). To generate this image, wild-type and DDR1 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-CD167a/DDR1 antibody [EPR24783-7] (ab288675) at 1/1000 dilution

Lane 1:

HeLa cell lysate at 20 µg

Lane 2:

NIH/3T3 cell lysate at 20 µg

Lane 3:

T-47D cell lysate at 20 µg

Lane 4:

Human Brain cell lysate at 20 µg

Lane 5:

Human Liver cell lysate at 20 µg

Lane 6:

Mouse Brain cell lysate at 20 µg

Lane 7:

Mouse Liver cell lysate at 20 µg

Lane 8:

Wild-type MCF7 Serum Starved (24 h) Vehicle Control Collagen I (0 ug/mL, 16 h) ab288242 cell lysate at 20 µg

Lane 9:

Wild-type MCF7 Serum Starved (24 h) Treated Collagen I (25 ug/mL, 16 h) ab288241 cell lysate at 20 µg

Lane 10:

DDR1 knockout MCF7 Serum Starved (24 h) Vehicle Control Collagen I (0 ug/mL, 16 h) ab288240 cell lysate at 20 µg

Lane 11:

DDR1 knockout MCF7 Serum Starved (24 h) Treated Collagen I (25 ug/mL, 16 h) ab288238 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

Predicted band size: 125 kDa

Observed band size: 100 kDa,125 kDa

false