Anti-USP9x antibody [EPR13809(B)] - BSA and Azide free

• WB

Lab

Western blot - Anti-USP9x antibody [EPR13809(B)] - BSA and Azide free (AB250191)

This data was developed using the same antibody clone in a different buffer formulation (ab180191).

Lanes 1- 2 : Merged signal (red and green). Green - ab180191 observed at 290 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.

ab180191 was shown to react with USP9x in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265665 (knockout cell lysate ab257790) was used. Wild-type HeLa and USP9X knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab180191 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-USP9x antibody [EPR13809(B)] - N-terminal (<a href='/en-us/products/primary-antibodies/usp9x-antibody-epr13809b-n-terminal-ab180191'>ab180191</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

USP9X knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human USP9X knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-usp9x-knockout-hela-cell-line-ab265665'>ab265665</a>)

Predicted band size: 140 kDa,292 kDa

Observed band size: 220 kDa,290 kDa

false

• IP

Lab

Immunoprecipitation - Anti-USP9x antibody [EPR13809(B)] - BSA and Azide free (AB250191)

ab180191 (purified) at 1/20 dilution immunoprecipitating ETS2 in HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 10 μg.
Lane 1 (input) : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 10 μg
Lane 2 (+) : ab180191 & HeLa whole cell lysate
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab180191 in HeLa whole cell lysate

For western blotting, ab180191 at 1/500 dilution (0.23 μg/mL) and veriBlot for IP secondary antibody (HRP) (ab131366) at 1/1000 dilution was used.

Blocking and diluting buffer : 5% NFDM /TBST.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab180191).

All lanes:

Immunoprecipitation - Anti-USP9x antibody [EPR13809(B)] - N-terminal (<a href='/en-us/products/primary-antibodies/usp9x-antibody-epr13809b-n-terminal-ab180191'>ab180191</a>)

Predicted band size: 292 kDa

false

• IP

Lab

Immunoprecipitation - Anti-USP9x antibody [EPR13809(B)] - BSA and Azide free (AB250191)

ab180191 (purified) at 1/20 dilution immunoprecipitating ETS2 in NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate 10 μg.

Lane 1 (input) : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate 10 μg.
Lane 2 (+) : ab180191 & NIH/3T3 whole cell lysate
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab180191 in NIH/3T3 whole cell lysate

For western blotting, ab180191 at 1/500 dilution (0.23 μg/mL) and veriBlot for IP secondary antibody (HRP) (ab131366) at 1/1000 dilution was used.

Blocking and diluting buffer : 5% NFDM /TBST.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab180191).

All lanes:

Immunoprecipitation - Anti-USP9x antibody [EPR13809(B)] - N-terminal (<a href='/en-us/products/primary-antibodies/usp9x-antibody-epr13809b-n-terminal-ab180191'>ab180191</a>)

Predicted band size: 292 kDa

false