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 – ab52971 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

ab52971 was shown to specifically react with Integrin beta 1 in wild-type HAP1 cells. No band was observed when Integrin beta 1 knockout samples were examined. Wild-type and Integrin beta 1 knockout samples were subjected to SDS-PAGE. ab52971 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 1 hour at room temperature before imaging.

ab52971 at 1/500 staining Integrin beta 1 antibody in human transitional cell carcinoma of bladder by immunohistochemistry (FFPE).

Immunohistochemical analysis of paraffin-embedded human transitional cell carcinoma of bladder tissue labeling Integrin beta 1 with ab52971 at 1/500 dilution followed by goat anti-rabbit IgG H&L (HRP) (ab97051, 1/500). Counter stained with hematoxylin.

Perform heat mediated antigen retrieval 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 - ab52971 observed at 140 kDa. Red signal from loading control – ab8245 observed at 37 kDa.

This western blot image is a comparison between ab52971 and a competitor's top cited rabbit polyclonal antibody.

All lanes : Anti-Integrin beta 1 antibody [EP1041Y] (ab52971) at 1/10000 dilution (purified)

Lane 1 : U87-MG cell lysate
Lane 2 : HT-1080 cell lysate
Lane 3 : U937 cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : HRP goat anti-rabbit (H+L) at 1/1000 dilution

Predicted band size: 88 kDa
Observed band size: 140 kDa why is the actual band size different from the predicted?

5% NFDM/TBST dilution buffer

All lanes : Anti-Integrin beta 1 antibody [EP1041Y] (ab52971) at 20 µg (unpurified)

Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : HT 1080 (Human fibrosarcoma) Whole Cell Lysate
Lane 3 : U2OS (Human osteosarcoma cell line) Whole Cell Lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (Alexa Fluor® 790) (ab175781) at 1/10000 dilution

Predicted band size: 88 kDa
Observed band size: 120 + 140 kDa why is the actual band size different from the predicted?

This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using Licor blocking buffer before being incubated with ab52971 overnight at 4°C. Antibody binding was detected using ab175781 at a 1:10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.

 

Secondary antibody - anti-rabbit Alexa Fluor 790

Unpurified ab52971 staining human breast cancer metastasis tissue sections by IHC-P. Sections were formaldehyde fixed and subjected to heat mediated antigen retrieval in citrate buffer (pH 6) prior to blocking with a commercial blocking reagent and incubation with the antibody (diluted 1/100) for 18 hours at 4°C.  A HRP-conjugated goat anti-rabbit was used as the secondary antibody.  This image shows a cancer metastasis at 40x with beta1 staining (in red) in both blood vessels and tumour cells. Blue is Hoechst for nuclei.  The antibody detection was enhanced using a commercial Cy3 tyramide signal amplification kit.

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Formaldehyde-fixed, paraffin-embedded human lung tissue stained for Integrin beta 1 using ab52971 at 1/100 dilution in immunohistochemical analysis.

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