Anti-EEF1A2 antibody [EPR22651-46] - BSA and Azide free

• IP

Unknown

Immunoprecipitation - Anti-EEF1A2 antibody [EPR22651-46] - BSA and Azide free (AB256523)

EEF1A2 was immunoprecipitated from 0.35 mg HEK-293T (human embryonic kidney epithelial cell) whole cell lysate with ab227824 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab227824 1/1000 dilution (0.54 μg/ml). VeriBlot for IP Detection Reagent (HRP) (ab131366) was used as the secondary antibody at 1/5000 dilution.

Lane 1 : HEK-293T (human embryonic kidney epithelial cell) whole cell lysate 10μg

Lane 2 : ab227824 IP in HEK-293T whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab227824 in HEK-293T whole cell lysate.

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

Exposure time : 15 seconds

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab227824).

All lanes:

Immunoprecipitation - Anti-EEF1A2 antibody [EPR22651-46] (<a href='/en-us/products/primary-antibodies/eef1a2-antibody-epr22651-46-ab227824'>ab227824</a>)

Predicted band size: 47 kDa,50 kDa

Observed band size: 46 kDa,50 kDa

false

• WB

Unknown

Western blot - Anti-EEF1A2 antibody [EPR22651-46] - BSA and Azide free (AB256523)

ab227824 was shown to specifically react with EEF1A2 in wild-type HAP1 cells as signal was lost in EEF1A2 knockout cells. Wild-type and EEF1A2 knockout samples were subjected to SDS-PAGE. ab227824 and ab181602 (Rabbit anti-GAPDH loading control) were incubated 1 hour at room temperature at 1/1000 dilution and 1/200,000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) secondary antibody at 1/100,000 dilution for 1 hour at room temperature before imaging. The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument using the ECL technique.

This antibody also recognizes a weakly unidentifiable band at around 75kDa

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

Exposure time : 3.25 seconds

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab227824).

All lanes:

Western blot - Anti-EEF1A2 antibody [EPR22651-46] (<a href='/en-us/products/primary-antibodies/eef1a2-antibody-epr22651-46-ab227824'>ab227824</a>) at 1/1000 dilution

Lane 1:

Wild-type HAP1 whole cell lysate at 20 µg

Lane 2:

EEF1A2 knockout HAP1 whole cell lysate at 20 µg

Lane 3:

HEK-293T (human embryonic kidney epithelial cell), whole cell lysate at 20 µg

Lane 4:

Neuro-2a (mouse neuroblastoma neuroblast), 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 1/100000 dilution

Predicted band size: 50 kDa

Observed band size: 50 kDa

false