Anti-MRas antibody [EPR12457] - BSA and Azide free

• IP

Lab

Immunoprecipitation - Anti-MRas antibody [EPR12457] - BSA and Azide free (AB240194)

This data was developed using ab176570, the same antibody clone in a different buffer formulation.
Purified ab176570 at 1/20 dilution (1μg) immunoprecipitating MRas in Human heart lysate.
Lane 1 (input) : Human heart lysate 10μg
Lane 2 (+) : ab176570 + Human heart lysate.
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab176570 in Human heart 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 : 24 kDa

All lanes:

Immunoprecipitation - Anti-MRas antibody [EPR12457] (<a href='/ko/products/primary-antibodies/mras-antibody-epr12457-ab176570'>ab176570</a>)

Predicted band size: 24 kDa

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• WB

Supplier Data

Western blot - Anti-MRas antibody [EPR12457] - BSA and Azide free (AB240194)

This data was developed using the same antibody clone in a different buffer formulation (ab176570).

All lanes:

Western blot - Anti-MRas antibody [EPR12457] (<a href='/ko/products/primary-antibodies/mras-antibody-epr12457-ab176570'>ab176570</a>) at 1/1000 dilution

All lanes:

Human brain lysate at 15 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 24 kDa

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• WB

Lab

Western blot - Anti-MRas antibody [EPR12457] - BSA and Azide free (AB240194)

This data was developed using the same antibody clone in a different buffer formulation (ab176570).

Lanes 1-3 : Merged signal (red and green). Green - ab176570 observed at 26 kDa. Red - loading control ab8245 observed at 36 kDa.

ab176570 Anti-MRas antibody [EPR12457] was shown to specifically react with MRas in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265510 (knockout cell lysate ab257541) was used. Wild-type and MRas knockout samples were subjected to SDS-PAGE. ab176570 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2.5 hours 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-MRas antibody [EPR12457] (<a href='/ko/products/primary-antibodies/mras-antibody-epr12457-ab176570'>ab176570</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

MRAS knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human MRAS knockout HeLa cell line (<a href='/ko/products/cell-lines/human-mras-knockout-hela-cell-line-ab265510'>ab265510</a>)

Lane 3:

Human brain tissue 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: 24 kDa

Observed band size: 26 kDa

false

• WB

Unknown

Western blot - Anti-MRas antibody [EPR12457] - BSA and Azide free (AB240194)

This data was developed using the same antibody clone in a different buffer formulation (ab176570).

All lanes:

Western blot - Anti-MRas antibody [EPR12457] (<a href='/ko/products/primary-antibodies/mras-antibody-epr12457-ab176570'>ab176570</a>) at 1/5000 dilution

Lane 1:

Mouse brain lysate at 15 µg

Lane 2:

Rat brain lysate at 15 µg

Secondary

All lanes:

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

Predicted band size: 24 kDa

false