Anti-DRAK2 antibody [EPR3163Y] - BSA and Azide free

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-DRAK2 antibody [EPR3163Y] - BSA and Azide free (AB201323)

This data was developed using ab108373, the same antibody clone in a different buffer formulation.

Immunofluorescence staining of Ramos cells with purified ab108373 at a working dilution of 1/100, counter-stained with DAPI. The secondary antibody was Alexa Fluor^® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor^® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab108373 was used at a dilution of 1/500 followed by an Alexa Fluor^® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor^® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.

• IP

Unknown

Immunoprecipitation - Anti-DRAK2 antibody [EPR3163Y] - BSA and Azide free (AB201323)

This data was developed using ab108373, the same antibody clone in a different buffer formulation.

ab108373 (purified) at 1/40 immunoprecipitating DRAK2 in 10 μg Ramos cell lysate (Lanes 1 and 2, observed at 42 kDa). Lane 3 - Rabbit monoclonal IgG (ab172730). For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution. Blocking buffer and concentration : 5% NFDM/TBST Dilution buffer and concentration : 5% NFDM/TBST

All lanes:

Immunoprecipitation - Anti-DRAK2 antibody [EPR3163Y] (<a href='/en-us/products/primary-antibodies/drak2-antibody-epr3163y-ab108373'>ab108373</a>)

Predicted band size: 39 kDa,42 kDa

false

• WB

Lab

Western blot - Anti-DRAK2 antibody [EPR3163Y] - BSA and Azide free (AB201323)

This data was developed using ab108373, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-DRAK2 antibody [EPR3163Y] (<a href='/en-us/products/primary-antibodies/drak2-antibody-epr3163y-ab108373'>ab108373</a>) at 1/10000 dilution

Lane 1:

Ramos whole cell lysate at 10 µg

Lane 2:

Raji whole cell lysate at 10 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: 42 kDa

Observed band size: 42 kDa

false

• WB

Lab

Western blot - Anti-DRAK2 antibody [EPR3163Y] - BSA and Azide free (AB201323)

This data was developed using ab108373, the same antibody clone in a different buffer formulation.

Lane 1 Wild-type HAP1 cell lysate (20 Âμg)

Lane 2 DRAK2 knockout HAP1 cell lysate (20 Âμg)

Lane 3 Ramos cell lysate (20 Âμg)

Lane 4 Raji cell lysate (20 Âμg)

Lanes 1 - 4 Merged signal (red and green). Green - ab108373 observed at 42 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab108373 was shown to recognize DRAK2 when DRAK2 knockout samples were used, along with additional cross-reactive bands. Wild-type and DRAK2 knockout samples were subjected to SDS-PAGE. ab108373 and ab8245 (loading control to GAPDH) were diluted 1/500 and 1/10000 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 1/10000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-DRAK2 antibody [EPR3163Y] (<a href='/en-us/products/primary-antibodies/drak2-antibody-epr3163y-ab108373'>ab108373</a>)

Predicted band size: 42 kDa

false

• WB

Unknown

Western blot - Anti-DRAK2 antibody [EPR3163Y] - BSA and Azide free (AB201323)

This data was developed using ab108373, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-DRAK2 antibody [EPR3163Y] (<a href='/en-us/products/primary-antibodies/drak2-antibody-epr3163y-ab108373'>ab108373</a>) at 1/500 dilution

Lane 1:

Jurkat cell lysate +TPA at 10 µg

Lane 2:

Ramos cell lysate at 10 µg

Lane 3:

Ramos cell lysate + TPA at 10 µg

Secondary

All lanes:

HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 42 kDa

false

• WB

Lab

Western blot - Anti-DRAK2 antibody [EPR3163Y] - BSA and Azide free (AB201323)

This data was developed using ab108373, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-DRAK2 antibody [EPR3163Y] (<a href='/en-us/products/primary-antibodies/drak2-antibody-epr3163y-ab108373'>ab108373</a>) at 1/1000 dilution

Lane 1:

C6 whole cell lysate at 10 µg

Lane 2:

PC-12 whole cell lysate at 10 µg

Lane 3:

NIH/3T3 whole cell lysate at 10 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: 42 kDa,59 kDa

Observed band size: 42 kDa

false