Anti-PP-T antibody [12F7]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-PP-T antibody [12F7] (AB223367)

4% formaldehyde-fixed HeLa (human epithelial cell line from cervix adnocarcinoma) cells stained for PP-T (green) using ab223367 at 1/100 dilution for 60 minutes at room temperature in ICC/IF, followed by Goat Anti-Mouse ATTO 488 at 1/100 dilution for 60 minutes at room temperature. Counterstain : DAPI (blue) nuclear stain at 1/5000 for 5 minutes room temperature.

(A) DAPI nuclear stain (B) ab223367 (C) Merge.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-PP-T antibody [12F7] (AB223367)

2% formaldehyde-fixed HEK-293 (human epithelial cell line from embryonic kidney) cells stained for PP-T (green) using ab223367 at 1/50 dilution in ICC/IF, followed by Goat Anti-Mouse Alexa Fluor 488 at 1/100 dilution for 60 minutes at RT. Counterstain : DAPI (blue) nuclear stain.

(A) DAPI nuclear stain (B) ab223367 (C) Merge.

• WB

Supplier Data

Western blot - Anti-PP-T antibody [12F7] (AB223367)

Blocking buffer : 5% skim milk.

Primary and secondary antibodies were incubated for 1 hour at room temperature.

All lanes:

Western blot - Anti-PP-T antibody [12F7] (ab223367) at 1/500 dilution

Lane 1:

A431 (human epidermoid carcinoma cell line) cell lysate at 15 µg

Lane 2:

HEK-293 (human epithelial cell line from embryonic kidney) cell lysate at 15 µg

Lane 3:

Jurkat (human T cell leukemia cell line from peripheral blood) cell lysate at 15 µg

Secondary

All lanes:

Goat Anti-Mouse IgG HRP at 1/200 dilution

Predicted band size: 56 kDa

Observed band size: 58 kDa

true

Exposure time: 6min

• WB

Lab

Western blot - Anti-PP-T antibody [12F7] (AB223367)

Lanes 1-4 : Merged signal (red and green). Green - ab223367 observed at 58 kDa. Red - loading control ab181602 observed at 36 kDa.

ab223367 Anti-PP-T antibody [12F7] was shown to specifically react with PP-T in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266507 (knockout cell lysate ab258136) was used. Wild-type and PP-T knockout samples were subjected to SDS-PAGE. ab223367 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preadsorbed (ab216777) and Goat anti-Mouse IgG H&L (IRDye^® 800CW) preadsorbed (ab216772) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-PP-T antibody [12F7] (ab223367) at 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

PPP5C knockout HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human PPP5C (PP-T) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-ppp5c-pp-t-knockout-hek-293t-cell-line-ab266507'>ab266507</a>)

Lane 3:

A431 cell lysate at 20 µg

Lane 4:

Jurkat cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-680rd-preadsorbed-ab216777'>ab216777</a>) at 1/10000 dilution

Predicted band size: 56 kDa

Observed band size: 58 kDa

false