Anti-USP14/TGT antibody [EPR15943] - BSA and Azide free

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-USP14/TGT antibody [EPR15943] - BSA and Azide free (AB251149)

This data was developed using ab192618, the same antibody clone in a different buffer formulation.

Intracellular flow cytometric analysis of Jurkat cells (2% paraformaldehyde-fixed) labeling USP14/TGT with ab192618 at 1/70 dilution (red) or a rabbit IgG (negative) (black), unlabeled cells (blue) followed by Goat anti rabbit IgG (FITC) secondary at 1/150 dilution.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-USP14/TGT antibody [EPR15943] - BSA and Azide free (AB251149)

This data was developed using ab192618, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of HeLa cells (4% Paraformaldehyde-fixed, 0.1% tritonX-100 permeabilized) labeling USP14/TGT with ab192618 at 1/70 dilution (5 μg/ml). A Goat anti rabbit IgG (Alexa Fluor488) at 1/400 dilution (ab150077) was used as secondary antibody. Counterstain : DAPI.

• WB

Lab

Western blot - Anti-USP14/TGT antibody [EPR15943] - BSA and Azide free (AB251149)

False colour image of Western blot : Anti-USP14/TGT antibody [EPR15943] - C-terminal staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab192618 was shown to bind specifically to USP14/TGT. A band was observed at 63 kDa in wild-type HeLa cell lysates with no signal observed at this size in USP14 CRISPR-Cas9 edited cell line ab266854 (CRISPR-Cas9 edited cell lysate ab257787). The band observed in the CRISPR-Cas9 edited lysate lane below 63 kDa is likely to represent a truncated form of USP14/TGT. 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 USP14 CRISPR-Cas9 edited HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % 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-USP14/TGT antibody [EPR15943] - C-terminal (<a href='/en-us/products/primary-antibodies/usp14-tgt-antibody-epr15943-c-terminal-ab192618'>ab192618</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

USP14 CRISPR-Cas9 edited HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human USP14 (TGT) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-usp14-tgt-knockout-hela-cell-line-ab266854'>ab266854</a>)

Lane 3:

USP14 knockout HeLa cell lysate at 20 µg

Predicted band size: 56 kDa

Observed band size: 63 kDa

false

• WB

Supplier Data

Western blot - Anti-USP14/TGT antibody [EPR15943] - BSA and Azide free (AB251149)

This data was developed using ab192618, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-USP14/TGT antibody [EPR15943] - C-terminal (<a href='/en-us/products/primary-antibodies/usp14-tgt-antibody-epr15943-c-terminal-ab192618'>ab192618</a>) at 1/2000 dilution

Lane 1:

HeLa cell lysate at 20 µg

Lane 2:

293T cell lysate at 20 µg

Lane 3:

Jurkat cell lysate at 20 µg

Lane 4:

HCT-116 cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugate at 1/1000 dilution

Predicted band size: 56 kDa

false

• WB

Supplier Data

Western blot - Anti-USP14/TGT antibody [EPR15943] - BSA and Azide free (AB251149)

This data was developed using ab192618, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-USP14/TGT antibody [EPR15943] - C-terminal (<a href='/en-us/products/primary-antibodies/usp14-tgt-antibody-epr15943-c-terminal-ab192618'>ab192618</a>) at 1/2000 dilution

Lane 1:

C6 cell lysate at 10 µg

Lane 2:

RAW 264.7 cell lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugate at 1/1000 dilution

Predicted band size: 56 kDa

false