Anti-USP14/TGT antibody [EPR15943] - C-terminal

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-USP14/TGT antibody [EPR15943] - C-terminal (AB192618)

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

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Immunocytochemistry/ Immunofluorescence - Anti-USP14/TGT antibody [EPR15943] - C-terminal (AB192618)

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] - C-terminal (AB192618)

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 knockout cell line ab266854 (knockout cell lysate ab257787). The band observed in the knockout 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 knockout 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 (ab192618) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lanes 2 - 3:

USP14 knockout HeLa cell lysate at 20 µg

Predicted band size: 56 kDa

Observed band size: 63 kDa

false

• WB

Lab

Western blot - Anti-USP14/TGT antibody [EPR15943] - C-terminal (AB192618)

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 (ab192618) 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] - C-terminal (AB192618)

All lanes:

Western blot - Anti-USP14/TGT antibody [EPR15943] - C-terminal (ab192618) 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] - C-terminal (AB192618)

All lanes:

Western blot - Anti-USP14/TGT antibody [EPR15943] - C-terminal (ab192618) 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