Anti-DNAJC9 antibody [EPR9856]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DNAJC9 antibody [EPR9856] (AB166612)

Immunohistochemical analysis of paraffin embedded Human tonsil tissue labeling DNAJC9 with ab166612 antibody at 1/100.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DNAJC9 antibody [EPR9856] (AB166612)

Immunohistochemical analysis of paraffin embedded Human ovarian carcinoma tissue labeling DNAJC9 with ab166612 antibody at 1/100.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• WB

Unknown

Western blot - Anti-DNAJC9 antibody [EPR9856] (AB166612)

All lanes:

Western blot - Anti-DNAJC9 antibody [EPR9856] (ab166612) 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 labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 30 kDa

Observed band size: 30 kDa

false

• IP

Unknown

Immunoprecipitation - Anti-DNAJC9 antibody [EPR9856] (AB166612)

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] (ab166612)

Predicted band size: 30 kDa

false

• WB

Lab

Western blot - Anti-DNAJC9 antibody [EPR9856] (AB166612)

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 knockout cell line ab266364 (knockout cell lysate ab257926). The band observed in the knockout 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 knockout 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] (ab166612) at 1/1000 dilution

Lane 1:

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

Lane 2:

DNAJC9 knockout HEK-293T cell lysate at 20 µg

Lane 3:

PC-3 cell lysate at 20 µg

Lane 4:

HepG2 cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/20000 dilution

Predicted band size: 30 kDa

false

• WB

Lab

Western blot - Anti-DNAJC9 antibody [EPR9856] (AB166612)

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] (ab166612) at 1/1000 dilution

Lane 1:

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

Lane 2:

DNAJC9 CRISPR-Cas9 edited HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human DNAJC9 knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-dnajc9-knockout-hek-293t-cell-line-ab266364'>ab266364</a>)

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

false