Anti-Integrin beta 3 antibody [EPR2342]

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Integrin beta 3 antibody [EPR2342] (AB119992)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized HUVEC (Human umbilical vein endothelial cell) cell line labeling Integrin beta 3 with ab119992 at 1/20 (red) compared with a Rabbit monoclonal IgG (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077), at 1/2000 dilution was used as the secondary antibody.

• IP

Supplier Data

Immunoprecipitation - Anti-Integrin beta 3 antibody [EPR2342] (AB119992)

Purified ab119992 at 1 : 20 dilution (0.7μg) immunoprecipitating Integrin beta 3 in HUVEC whole cell lysate.

Lane 1 (input) : HUVEC (Human umbilical vein endothelial cell) whole cell lysate 10μg.
Lane 2 (+) : ab119992 + HUVEC whole cell lysate.
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab119992 in HUVEC whole cell lysate.
VeriBlot for IP Detection Reagent (HRP)(ab131366) (1 : 1000 dilution) was used for Western blotting.

Blocking Buffer and concentration : 5% NFDM/TBST.
Diluting buffer and concentration : 5% NFDM/TBST.
Observed band size : 110 kDa

All lanes:

Immunoprecipitation - Anti-Integrin beta 3 antibody [EPR2342] (ab119992)

Predicted band size: 87 kDa

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• WB

Lab

Western blot - Anti-Integrin beta 3 antibody [EPR2342] (AB119992)

Anti-Integrin beta antibody [EPR20825] (ab210515) at 1/1000 dilution or Anti-Integrin beta 3 antibody [EPR2342] (ab119992) at 1/500.

Comparison of staining with an antibody specific to Integrin beta 3 (ab119992).

The Integrin beta 3 immunogen used for this product shares the following sequence identities with other Integrin beta proteins : beta 5 – 61%, beta 6 – 53%, beta 7 – 48%, beta 1 – 47%, beta 2 – 46%, beta 8 – 42%, beta 4 – 38%. Specific cross-reactivities have not been determined experimentally, but based on our data we expect that the antibody cross-reacts with 1 or more of these proteins.

Exposure :

For ab210515 : 1, 2, 5 lanes : 3 s; 3, 4 lanes : 37 s; 6 lane : 3 min; 7 lane : 15 s; 8 lane : 114 s.

For ab119992 : 1, 2 lane : 10 s; 3~8 lanes : 3 min

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Lanes 1 - 8:

Western blot - Anti-Integrin beta antibody [EPR20825] (<a href='/en-us/products/primary-antibodies/integrin-beta-antibody-epr20825-ab210515'>ab210515</a>) at 1/1000 dilution

Lanes 1 - 8:

Western blot - Anti-Integrin beta 3 antibody [EPR2342] (ab119992) at 1/500 dilution

Lane 1:

U-87MG (Human glioblastoma-astrocytoma epithelial cell) whole cell lysates at 20 µg

Lane 2:

HEL (Human Erythroleukemia erythroblast) whole cell lysates at 20 µg

Lane 3:

LnCaP (Human prostate carcinoma epithelial cell) whole cell lysates at 20 µg

Lane 4:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates at 20 µg

Lane 5:

HUVEC (Human umbilical vein endothelial cell) whole cell lysates at 20 µg

Lane 6:

PC-3 (Human prostate adenocarcinoma epithelial cell) whole cell lysates at 20 µg

Lane 7:

C2C12 (Mouse myoblasts myoblast) whole cell lysates at 20 µg

Lane 8:

Raw264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysates at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 87 kDa

Observed band size: 110 kDa

false

• WB

Lab

Western blot - Anti-Integrin beta 3 antibody [EPR2342] (AB119992)

Blocking buffer and concentration : 5% NFDM/TBST. Diluting buffer and concentration : 5% NFDM/TBST. ab181602 was used as a loading control. LnCaP, HeLa and PC-3 are low expression cell lines ( PMID : 21867506, PMID : 25248000). Though there are publications stating expression of Integrin beta 3 in RAW264.7, ab119992 and ab179473 show negative in Raw264.7.

All lanes:

Western blot - Anti-Integrin beta 3 antibody [EPR2342] (ab119992) at 1/1000 dilution

Lane 1:

U-87MG (Human glioblastoma-astrocytoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

HEL (Human Erythroleukemia erythroblast) whole cell lysate at 20 µg

Lane 3:

LnCaP (Human prostate carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 5:

HUVEC (Human umbilical vein endothelial cell) whole cell lysate at 20 µg

Lane 6:

PC-3 (Human prostate adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 7:

C2C12 (Mouse myoblasts myoblast) whole cell lysate at 20 µg

Lane 8:

Raw264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 87 kDa

Observed band size: 110 kDa

false

Exposure time: 40s