Anti-TRIM24 antibody [EPR22825-2]

9 product images

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  Western blot - Anti-TRIM24 antibody [EPR22825-2] (ab256491)

  Lanes 1- 4: Merged signal (red and green). Green - ab256491 observed at 140 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) observed at 37 kDa.
  

  ab256491 was shown to react with Tripartite Motif Containing 24 in wild-type HeLa cells in western blot. The band observed in knockout cell line Human TRIM24 knockout HeLa cell line ab264963 (knockout cell lysate Human TRIM24 knockout HeLa cell lysate ab258246) lane below 140kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HeLa and TRIM24 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab256491 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^®800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^®680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  All lanes: Western blot - Anti-TRIM24 antibody [EPR22825-2] (ab256491) at 1/1000 dilution

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

  Lane 2: TRIM24 knockout HeLa cell lysate at 20 µg

  Lane 3: Hap1 cell lysate at 20 µg

  Lane 4: A549 cell lysate at 20 µg

  Performed under reducing conditions.

  Predicted band size: 116 kDa

  Observed band size: 140 kDa

• 

  Western blot - Anti-TRIM24 antibody [EPR22825-2] (ab256491)

  The expression profile observed is consistent with what has been described in the literature (PMID: 19909775).
  

  ab256491 was shown to specifically react with TRIM24 in wild-type HAP1 cells as signal was lost in TRIM24 knockout cells. Wild-type and TRIM24 knockout samples were subjected to SDS-PAGE. ab256491 and Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 (Rabbit anti-GAPDH loading control) were incubated 1 hour at room temperature at 1/1000 dilution and 1/200,000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) secondary antibody at 1/100,000 dilution for 1 hour at room temperature before imaging. The blot was developed on a BIO-RAD^® ChemiDoc™ MP instrument using the ECL technique. Blocking/Diluting buffer and concentration: 5% NFDM/TBST. Exposure Time: 48 seconds.

  All lanes: Western blot - Anti-TRIM24 antibody [EPR22825-2] (ab256491) at 1/1000 dilution

  Lane 1: Wild-type HAP1 whole cell lysate at 20 µg

  Lane 2: TRIM24 knockout HAP1 whole cell lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Predicted band size: 116 kDa

  Observed band size: 140 kDa

  The expression profile observed is consistent with what has been described in the literature (PMID: 19909775).
  

  ab256491 was shown to specifically react with TRIM24 in wild-type HAP1 cells as signal was lost in TRIM24 knockout cells. Wild-type and TRIM24 knockout samples were subjected to SDS-PAGE. ab256491 and Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 (Rabbit anti-GAPDH loading control) were incubated 1 hour at room temperature at 1/1000 dilution and 1/200,000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) secondary antibody at 1/100,000 dilution for 1 hour at room temperature before imaging. The blot was developed on a BIO-RAD^® ChemiDoc™ MP instrument using the ECL technique. Blocking/Diluting buffer and concentration: 5% NFDM/TBST. Exposure Time: 48 seconds.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab256491).

• 

  Western blot - Anti-TRIM24 antibody [EPR22825-2] (ab256491)

  Lanes 1- 2: Merged signal (red and green). Green - Anti-NFkB p100/NFKB2 antibody [EPR4686-66] ab175192 observed at 120 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) observed at 37 kDa.
  

  Anti-NFkB p100/NFKB2 antibody [EPR4686-66] ab175192 was shown to react with NFkB p100/NFKB2 in wild-type HCT116 cells in western blot. The band observed in knockout cell line Human NFKB2 (NFkB p100/NFKB2) knockout HCT116 cell line ab266883 (knockout cell lysate Human NFKB2 (NFkB p100/NFKB2) knockout HCT116 cell lysate ab257245) lane below 97kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HCT116 and NFKB2 knockout HCT117 cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. Anti-NFkB p100/NFKB2 antibody [EPR4686-66] ab175192 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated overnight at 4°C at a 1 in 10000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^®800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^®680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  Lane 1: Western blot - Anti-NFkB p100/NFKB2 antibody [EPR4686-66] (Anti-NFkB p100/NFKB2 antibody [EPR4686-66] ab175192) at 1/1000 dilution

  Lane 2: Western blot - Anti-TRIM24 antibody [EPR22825-2] (ab256491) at 1/1000 dilution

  Lane 1: Wild-type HCT116 cell lysate at 20 µg

  Lane 2: NFKB2 knockout HCT116 cell lysate at 20 µg

  Performed under reducing conditions.

  Predicted band size: 97 kDa

  Observed band size: 120 kDa

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRIM24 antibody [EPR22825-2] (ab256491)

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue labeling TRIM24 with ab256491 at 1/500 dilution (1.08μg/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). Nuclear staining in human breast cancer (PMID: 21164480) is observed. The section was incubated with Anti-BAP1 antibody [EPR22826-65] ab255611 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin.
  

  Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).

  Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

• 

  Western blot - Anti-TRIM24 antibody [EPR22825-2] (ab256491)

  The expression profile observed is consistent with what has been described in the literature (PMID: 19909775). Blocking/Diluting buffer and concentration: 5% NFDM/TBST. Exposure Time: Lane 1: 3 minutes. Lanes 2-5: 15 seconds.

  All lanes: Western blot - Anti-TRIM24 antibody [EPR22825-2] (ab256491) at 1/1000 dilution

  Lane 1: WEHI-3 (mouse leukemia lymphoblast), whole cell lysate at 20 µg

  Lane 2: P815 (mouse mastocytoma mast Cell), whole cell lysate at 20 µg

  Lane 3: CTLL-2 (mouse T lymphocyte), whole cell lysate at 20 µg

  Lane 4: C2C12 (mouse myoblasts myoblast), whole cell lysate at 20 µg

  Lane 5: Mouse colon tissue lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Predicted band size: 116 kDa

  Observed band size: 140 kDa

• 

  Western blot - Anti-TRIM24 antibody [EPR22825-2] (ab256491)

  The expression profile observed is consistent with what has been described in the literature (PMID: 19909775). Blocking/Diluting buffer and concentration: 5% NFDM/TBST. Exposure Time: 48 seconds.

  All lanes: Western blot - Anti-TRIM24 antibody [EPR22825-2] (ab256491) at 1/1000 dilution

  Lane 1: HCT116 (human colorectal carcinoma epithelial cell), whole cell lysate at 20 µg

  Lane 2: HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate at 20 µg

  Lane 3: PC-3 (human prostate adenocarcinoma epithelial cell), whole cell lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Predicted band size: 116 kDa

  Observed band size: 140 kDa

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRIM24 antibody [EPR22825-2] (ab256491)

  Immunohistochemical analysis of paraffin-embedded human lung cancer tissue labeling TRIM24 with ab256491 at 1/500 dilution (1.08μg/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). Nuclear staining in human lung cancer (PMID: 22666376) is observed. The section was incubated with Anti-BAP1 antibody [EPR22826-65] ab255611 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin.
  

  Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).

  Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

• 

  Immunocytochemistry/ Immunofluorescence - Anti-TRIM24 antibody [EPR22825-2] (ab256491)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Wild type HAP1 (Wild type human chronic myelogenous leukemia near-haploid cell line) and TRIM24 knockout HAP1 (TRIM24 knockout human chronic myelogenous leukemia near-haploid cell line) cells labeling TRIM24 with ab256491 at 1/100 5.4 μg/ml dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary antibody at 1/1000 2 μg/ml dilution (Green). Confocal image showing nuclear staining in Wild type HAP1 cell line, and no staining in TRIM24 knockout HAP1 cell line. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (Red). The nuclear counterstain was DAPI (Blue).
  

  Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary at 1/1000 2 μg/ml dilution.

• 

  Immunocytochemistry/ Immunofluorescence - Anti-TRIM24 antibody [EPR22825-2] (ab256491)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HepG2 (human hepatocellular carcinoma epithelial cell) cells labeling TRIM24 with ab256491 at 1/100 5.4 μg/ml dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary antibody at 1/1000 2 μg/ml dilution (Green). Confocal image showing nuclear staining in HepG2 cell line is observed. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (Red). The nuclear counterstain was DAPI (Blue).
  

  Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary at 1/1000 2 μg/ml dilution.