Lanes 1, 5 and 9: Wild-type HAP1 cell lysate (20 µg)
Lanes 2, 6 and 10: Integrin beta 1 knockout HAP1 cell lysate (20 µg)
Lanes 3, 7 and 11: U87-MG cell lysate (20 µg)
Lanes 4, 8 and 12: A431 cell lysate (20 µg)
Lanes 1, 2, 3 and 4: Green signal from target – ab179471 observed at 140 kDa
Lanes 5, 6, 7 and 8: Red signal from loading control – ab8245 observed at 37 kDa
Lanes 9, 10, 11 and 12: Merged (red and green) signal

ab179471 was shown to react with Integrin beta 1 in wild-type HAP1 cells as well as additional cross-reactive bands. No bands were observed when Integrin beta 1 knockout samples were examined. Wild-type and Integrin beta 1 knockout samples were subjected to SDS-PAGE. ab179471 and ab8245 (loading control to GAPDH) were diluted 1/10,000 and 1/2000 respectively and incubated overnight at 4ºC. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10,000 dilution for 1hr at room temperature before imaging.

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Integrin beta 1 with ab179471 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Membrane staining on epithelial cells of Human colon is observed. Counter stained with Hematoxylin.

Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Immunohistochemical analysis of paraffin-embedded Human hepatocellular carcinoma tissue labeling Integrin beta 1 with ab179471 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Membrane staining on tumor cells of Human hepatocellular carcinoma is observed. Counter stained with Hematoxylin.

Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: Integrin beta 1 knockout HAP1 cell lysate (20 µg)
Lane 3: U87-MG cell lysate (20 µg)
Lane 4: A431 cell lysate (20 µg)

Lanes 1 - 4: Merged signal (red and green). Green - ab179471 observed at 140 kDa. Red signal from loading control – ab8245 observed at 37 kDa.

This western blot image is a comparison between ab179471 and a competitor's top cited rabbit polyclonal antibody.

Immunohistochemical analysis of paraffin-embedded Rat stomach tissue labeling Integrin beta 1 with ab179471 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Membrane staining on epithelial cells of rat stomach is observed. Counter stained with Hematoxylin.

Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling Integrin beta 1 with ab179471 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Membrane and cytoplasmic staining on mouse kidney is observed. Counter stained with Hematoxylin.

Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Anti-Integrin beta 1 antibody [EPR16895] (ab179471) at 1/20000 dilution + A431 (Human epidermoid carcinoma) whole cell lysates at 10 µg

Secondary
Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 88 kDa
Observed band size: 130-150 kDa why is the actual band size different from the predicted?

Blocking/Dilution buffer: 5% NFDM/TBST.

Anti-Integrin beta 1 antibody [EPR16895] (ab179471) at 1/20000 dilution + U-87 MG (Human glioblastoma-astrocytoma epithelial cell line) whole cell lysates at 10 µg

Secondary
Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 88 kDa
Observed band size: 130-150 kDa why is the actual band size different from the predicted?

Blocking/Dilution buffer: 5% NFDM/TBST.

Exposure time = 1 minute

Anti-Integrin beta 1 antibody [EPR16895] (ab179471) at 1/2000 dilution + Human spleen lysates at 10 µg

Secondary
Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

Predicted band size: 88 kDa
Observed band size: 130-150 kDa why is the actual band size different from the predicted?

Blocking/Dilution buffer: 5% NFDM/TBST.

Exposure time = 1 minute

All lanes : Anti-Integrin beta 1 antibody [EPR16895] (ab179471) at 1/2000 dilution

Lane 1 : Mouse heart lysates
Lane 2 : Mouse kidney lysates
Lane 3 : Rat heart lysates
Lane 4 : Rat kidney lysates
Lane 5 : Rat spleen lysates

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 88 kDa
Observed band size: 130-150 kDa why is the actual band size different from the predicted?


Exposure time: 1 minute

Blocking/Dilution buffer: 5% NFDM/TBST.