Anti-BAG2 antibody [EPR3567] - BSA and Azide free

• IP

Lab

Immunoprecipitation - Anti-BAG2 antibody [EPR3567] - BSA and Azide free (AB247500)

This data was developed using ab79406, the same antibody clone in a different buffer formulation.

BAG2 was immunoprecipitated from 0.35 mg Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate 10 μg with 79406 at 1/50 dilution (2μg). VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate 10 μg

Lane 2 : ab79406 IP in Jurkat whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab79406 in Jurkat whole cell lysate

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

All lanes:

Immunoprecipitation - Anti-BAG2 antibody [EPR3567] (<a href='/ko/products/primary-antibodies/bag2-antibody-epr3567-ab79406'>ab79406</a>)

Predicted band size: 23 kDa

Observed band size: 24 kDa

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• WB

Lab

Western blot - Anti-BAG2 antibody [EPR3567] - BSA and Azide free (AB247500)

This data was developed using the same antibody clone in a different buffer formulation (ab79406).

Lanes 1-3 : Merged signal (red and green). Green - ab79406 observed at 25 kDa. Red - loading control ab8245 observed at 36 kDa.

ab79406 Anti-BAG2 antibody [EPR3567] was shown to specifically react with BAG2 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265907 (knockout cell lysate ab257369) was used. Wild-type and BAG2 knockout samples were subjected to SDS-PAGE. ab79406 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. 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 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-BAG2 antibody [EPR3567] (<a href='/ko/products/primary-antibodies/bag2-antibody-epr3567-ab79406'>ab79406</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

BAG2 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human BAG2 knockout HeLa cell line (<a href='/ko/products/cell-lines/human-bag2-knockout-hela-cell-line-ab265907'>ab265907</a>)

Lane 3:

HepG2 cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 23 kDa

Observed band size: 25 kDa

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