Anti-HIP1 antibody [EPR10814] - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-HIP1 antibody [EPR10814] - BSA and Azide free (AB250453)

This data was developed using ab181238, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of HeLa cells (-20°C Acetone-fixed) labeling HIP1 with ab181238 at 1/250 dilution, followed by Goat anti rabbit IgG (Alexa Fluor® 488) secondary at 1/200 dilution and counter-stained with DAPI (blue).

• IP

Supplier Data

Immunoprecipitation - Anti-HIP1 antibody [EPR10814] - BSA and Azide free (AB250453)

This data was developed using ab181238, the same antibody clone in a different buffer formulation.

Western blot analysis of immunoprecipitation pellet from A549 cell lysate immunoprecipitated using ab181238 at 1/50 dilution.
Secondary : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution.

All lanes:

Immunoprecipitation - Anti-HIP1 antibody [EPR10814] (<a href='/en-us/products/primary-antibodies/hip1-antibody-epr10814-ab181238'>ab181238</a>)

Predicted band size: 116 kDa

false

• WB

Lab

Western blot - Anti-HIP1 antibody [EPR10814] - BSA and Azide free (AB250453)

This data was developed using the same antibody clone in a different buffer formulation (ab181238).

Lanes 1 - 3 : Merged signal (red and green). Green - ab181238 observed at 116 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

ab181238 was shown to react with HIP1 in wild-type HAP1 cells in western blot with loss of signal observed in HIP1 knockout sample. Wild-type and HIP1 knockout HAP1 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween^®) before incubation with ab181238 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 1000 Dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-HIP1 antibody [EPR10814] (<a href='/en-us/products/primary-antibodies/hip1-antibody-epr10814-ab181238'>ab181238</a>) at 1/1000 dilution

Lane 1:

wild-type HAP1 cell lysate at 20 µg

Lane 2:

HIP1 knockout HAP1 cell lysate at 20 µg

Lane 3:

HeLa cell lysate at 20 µg

Lane 4:

Jurkat cell lysate at 20 µg

Predicted band size: 116 kDa

Observed band size: 116 kDa

false

• WB

Supplier Data

Western blot - Anti-HIP1 antibody [EPR10814] - BSA and Azide free (AB250453)

This data was developed using ab181238, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-HIP1 antibody [EPR10814] (<a href='/en-us/products/primary-antibodies/hip1-antibody-epr10814-ab181238'>ab181238</a>) at 1/2000 dilution

Lane 1:

HCT-116 cell lysate at 20 µg

Lane 2:

HeLa cell lysate at 20 µg

Lane 3:

Jurkat cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 116 kDa

false