Anti-DNAJC9 antibody [EPR9856] - BSA and Azide free

• IP

Unknown

Immunoprecipitation - Anti-DNAJC9 antibody [EPR9856] - BSA and Azide free (AB249352)

This data was developed using ab166612, the same antibody clone in a different buffer formulation.Immunoprecipitation. ab166612 at 1/1000 staining DNAJC9 in 293T cell lysate immunoprecipitated using ab166612 at 1/10. Predicted band size : 30 kDa.

All lanes:

Immunoprecipitation - Anti-DNAJC9 antibody [EPR9856] - BSA and Azide free (ab249352)

Predicted band size: 30 kDa

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• WB

Lab

Western blot - Anti-DNAJC9 antibody [EPR9856] - BSA and Azide free (AB249352)

False colour image of Western blot : Anti-DNAJC9 antibody [EPR9856] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab166612 was shown to bind specifically to DNAJC9. A band was observed at 35 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in DNAJC9 CRISPR-Cas9 edited cell line ab266364 (CRISPR-Cas9 edited cell lysate ab257926). The band observed in the CRISPR-Cas9 edited lysate lane below 35 kDa is likely to represent a truncated form of DNAJC9. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and DNAJC9 CRISPR-Cas9 edited HEK-293T 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 then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

All lanes:

Western blot - Anti-DNAJC9 antibody [EPR9856] (<a href='/en-us/products/primary-antibodies/dnajc9-antibody-epr9856-ab166612'>ab166612</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human DNAJC9 knockout HEK-293T cell lysate (<a href='/en-us/products/cell-lysates/human-dnajc9-knockout-hek-293t-cell-lysate-ab257926'>ab257926</a>) at 20 µg

Lane 3:

PC-3 cell lysate at 20 µg

Lane 4:

HepG2 cell lysate at 20 µg

Predicted band size: 30 kDa

Observed band size: 35 kDa

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• WB

Unknown

Western blot - Anti-DNAJC9 antibody [EPR9856] - BSA and Azide free (AB249352)

This data was developed using ab166612, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-DNAJC9 antibody [EPR9856] - BSA and Azide free (ab249352) at 1/1000 dilution

Lane 1:

HepG2 cell lysate at 10 µg

Lane 2:

U87-MG cell lysate at 10 µg

Lane 3:

Raji cell lysate at 10 µg

Lane 4:

SH-SY5Y cell lysate at 10 µg

Lane 5:

293T cell lysate at 10 µg

Lane 6:

Jurkat cell lysate at 10 µg

Secondary

All lanes:

HRP labeled goat anti-rabbit at 1/2000 dilution

Predicted band size: 30 kDa

Observed band size: 30 kDa

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