Anti-HAUSP / USP7 antibody [EPR4253] - BSA and Azide free

• WB

Lab

Western blot - Anti-HAUSP / USP7 antibody [EPR4253] - BSA and Azide free (AB239936)

This data was developed using ab108931, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) 1 : 1000,000 dilution.

Exposure time : Lane 1 : 60 seconds; Lane 2 : 3 seconds

All lanes:

Western blot - Anti-HAUSP / USP7 antibody [EPR4253] (<a href='/en-us/products/primary-antibodies/hausp-usp7-antibody-epr4253-ab108931'>ab108931</a>) at 1/1000 dilution

Lane 1:

Rat spinal cord tissue lysate at 20 µg

Lane 2:

PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 128 kDa

false

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-HAUSP / USP7 antibody [EPR4253] - BSA and Azide free (AB239936)

ab108931, at 1/100, staining HAUSP / USP7 in HeLa cells by Immunofluorescence.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108931).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HAUSP / USP7 antibody [EPR4253] - BSA and Azide free (AB239936)

ab108931, at 1/50, staining HAUSP / USP7 in Human colon tissue by Immunohistochemistry, Paraffin-embedded tissue.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108931).

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-HAUSP / USP7 antibody [EPR4253] - BSA and Azide free (AB239936)

Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling HAUSP / USP7 with unpurified ab108931 at 1/30 dilution (10ug/ml) (red). Cells were fixed with 80% methanol and permeabilised with 0.1% Tween-20. A Goat anti rabbit IgG (Alexa Fluor^® 488) (ab150077) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) (ab172730) was used as the isotype control, Cell without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108931).

• WB

Lab

Western blot - Anti-HAUSP / USP7 antibody [EPR4253] - BSA and Azide free (AB239936)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108931).

Lanes 1- 2 : Merged signal (red and green). Green - ab108931 observed at 128 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab108931 was shown to react with HAUSP / USP7 in wild-type HEK-293T cells in western blot. The band observed in CRISPR/Cas9 edited cell line ab266535 (CRISPR/Cas9 edited cell lysate ab257284) lane below 128kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HEK-293T and USP7 CRISPR/Cas9 edited HEK-293T 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. ab108931 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) 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-HAUSP / USP7 antibody [EPR4253] (<a href='/en-us/products/primary-antibodies/hausp-usp7-antibody-epr4253-ab108931'>ab108931</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

USP7 CRISPR/Cas9 edited HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human USP7 (HAUSP) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-usp7-hausp-knockout-hek-293t-cell-line-ab266535'>ab266535</a>)

Predicted band size: 128 kDa

Observed band size: 128 kDa

false

• WB

Lab

Western blot - Anti-HAUSP / USP7 antibody [EPR4253] - BSA and Azide free (AB239936)

This data was developed using the same antibody clone in a different buffer formulation (ab108931).

Lane 1 : Wild-type HAP1 cell lysate (20 μg)
Lane 2 : HAUSP/USP7 knockout HAP1 cell lysate (20 μg)
Lane 3 : HeLa cell lysate (20 μg)
Lane 4 : MCF7 cell lysate (20 μg)
Lanes 1 - 4 : Merged signal (red and green). Green - ab108931 observed at 115 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab108931 was shown to specifically react with HAUSP/USP7 when HAUSP/USP7 knockout samples were used. Wild-type and HAUSP/USP7 knockout samples were subjected to SDS-PAGE. ab108931 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/2000 respectively and incubated overnight at 4°C. 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 1/10 000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-HAUSP / USP7 antibody [EPR4253] (<a href='/en-us/products/primary-antibodies/hausp-usp7-antibody-epr4253-ab108931'>ab108931</a>)

Predicted band size: 128 kDa

false