Anti-Integrin beta 1 antibody [EP1041Y] - BSA and Azide free

• IHC-P

AbReview9866****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Integrin beta 1 antibody [EP1041Y] - BSA and Azide free (AB192456)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52971).

This image is courtesy of an anonymous Abreview

• IHC-P

AbReview13423****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Integrin beta 1 antibody [EP1041Y] - BSA and Azide free (AB192456)

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52971).

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

This image is courtesy of an Abreview submitted by Mark Southwood.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Integrin beta 1 antibody [EP1041Y] - BSA and Azide free (AB192456)

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. This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52971). Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• WB

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Western blot - Anti-Integrin beta 1 antibody [EP1041Y] - BSA and Azide free (AB192456)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52971).

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 4°C. 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.

All lanes:

Western blot - Anti-Integrin beta 1 antibody [EP1041Y] - BSA and Azide free (ab192456)

Predicted band size: 88 kDa

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• WB

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Western blot - Anti-Integrin beta 1 antibody [EP1041Y] - BSA and Azide free (AB192456)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52971).

5% NFDM/TBST dilution buffer

All lanes:

Western blot - Anti-Integrin beta 1 antibody [EP1041Y] - BSA and Azide free (ab192456) at 1/10000 dilution

Lane 1:

U87-MG cell lysate at 20 µg

Lane 2:

HT-1080 cell lysate at 20 µg

Lane 3:

U937 cell lysate at 20 µg

Secondary

All lanes:

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

Predicted band size: 88 kDa

false

• WB

Unknown

Western blot - Anti-Integrin beta 1 antibody [EP1041Y] - BSA and Azide free (AB192456)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52971).

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 (ab175781)

All lanes:

Western blot - Anti-Integrin beta 1 antibody [EP1041Y] - BSA and Azide free (ab192456) at 20 µg

Lane 1:

HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 20 µg

Lane 2:

HT 1080 (Human fibrosarcoma) Whole Cell Lysate at 20 µg

Lane 3:

U2OS (Human osteosarcoma cell line) Whole Cell Lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 790) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-alexa-fluor-790-ab175781'>ab175781</a>) at 1/10000 dilution

Predicted band size: 88 kDa

Observed band size: 120 kDa,140 kDa

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• WB

Unknown

Western blot - Anti-Integrin beta 1 antibody [EP1041Y] - BSA and Azide free (AB192456)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52971).

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:

Western blot - Anti-Integrin beta 1 antibody [EP1041Y] - BSA and Azide free (ab192456)

Predicted band size: 88 kDa

false