Anti-HOXA9 antibody [EPR3655(2)] - BSA and Azide free

• IP

Supplier Data

Immunoprecipitation - Anti-HOXA9 antibody [EPR3655(2)] - BSA and Azide free (AB248915)

Lane 1 : Wild-type HEK-293 input Lane 2 : HOXA9 knockout HEK-293 input Lane 3 : Wild-type HEK-293 immunoprecipitated with anti-HOXA9 antibody ab140631 Lane 4 : HOXA9 knockout HEK-293 immunoprecipitated with anti-HOXA9 antibody ab140631 Lane 5 : Wild-type HEK-293 immunoprecipitated with isotype control antibody ab172730 Lane 6 : HOXA9 knockout HEK-293 immunoprecipitated with isotype control antibody ab172730 HOXA9 was immunoprecipitated with Recombinant Anti-HOXA9 antibody [EPR3655(2)] (ab140631) at 10 µg ml-1. A non-specific band detected in the input material at 37 kDa was not immunoprecipitated. ab172730 was used as the isotype control. Western blot was performed on immunoprecipitates of wild-type and HOXA9 knockout HEK-293 cell lysates (ab273706). To generate this image, wild-type and HOXA9 knockout HEK-293 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with VeriBlot for IP Detection Reagent (HRP) (ab131366) for 1 h at room temperature, washed again four times then imaged. This data was developed using the same antibody clone in a different buffer formulation (ab140631).

All lanes:

Immunoprecipitation - Anti-HOXA9 antibody [EPR3655(2)] (<a href='/en-us/products/primary-antibodies/hoxa9-antibody-epr36552-ab140631'>ab140631</a>) at 10 µg/mL

Lane 1:

Wild-type HEK-293 input

Lane 2:

HOXA9 knockout HEK-293 input

Lane 3:

Wild-type HEK-293 immunoprecipitated with anti-HOXA9 antibody <a href='/en-us/products/primary-antibodies/hoxa9-antibody-epr36552-ab140631'>ab140631</a>

Lane 4:

HOXA9 knockout HEK-293 immunoprecipitated with anti-HOXA9 antibody <a href='/en-us/products/primary-antibodies/hoxa9-antibody-epr36552-ab140631'>ab140631</a>

Lane 5:

Wild-type HEK-293 immunoprecipitated with isotype control antibody <a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>

Lane 6:

HOXA9 knockout HEK-293 immunoprecipitated with isotype control antibody <a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>)

Predicted band size: 30 kDa

false

• WB

Lab

Western blot - Anti-HOXA9 antibody [EPR3655(2)] - BSA and Azide free (AB248915)

This data was developed using ab140631, the same antibody clone in a different buffer formulation.

Western blot : Anti-HOXA9 antibody [EPR3655(2)] (ab140631) staining at 1/1000 dilution, shown in black; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab140631 was shown to bind specifically to HOXA9. A band was observed at 30 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in HOXA9 knockout cell line. To generate this image, wild-type and HOXA9 knockout HCT 116 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times before development with a high-sensitivity ECL substrate kit and imaged with 1 minute exposure time. Secondary antibodies used were HRP conjugated Goat anti-Rabbit (H+L) and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-HOXA9 antibody [EPR3655(2)] (<a href='/en-us/products/primary-antibodies/hoxa9-antibody-epr36552-ab140631'>ab140631</a>) at 1/1000 dilution

Lane 1:

Wild-type HCT 116 cell lysate at 20 µg

Lane 2:

Western blot - Human HOXA9 knockout HCT116 cell line (<a href='/en-us/products/cell-lines/human-hoxa9-knockout-hct116-cell-line-ab287488'>ab287488</a>)

Lane 2:

HOXA9 knockout HCT 116 cell lysate at 20 µg

Secondary

Lanes 1 - 2:

HRP conjugated Goat anti-Rabbit (H+L) at 1/20000 dilution

Lanes 1 - 2:

Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Observed band size: 30 kDa

false