Anti-USP22 antibody [EPR18945] - BSA and Azide free

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-USP22 antibody [EPR18945] - BSA and Azide free (AB238948)

Immunohistochemical analysis of paraffin-embedded Human breast carcinoma tissue labeling USP22 with ab195289 at 1/100 followed by a Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) Ready to use. Nuclear staining on human breast carcinoma. The section was incubated with ab195289 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) Ready to use.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab195289).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-USP22 antibody [EPR18945] - BSA and Azide free (AB238948)

Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling USP22 with ab195289 at 1/100 followed by a Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) Ready to use dilution. Nuclear staining on human cerebrum. The section was incubated with ab195289 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) Ready to use.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab195289).

• IP

Supplier Data

Immunoprecipitation - Anti-USP22 antibody [EPR18945] - BSA and Azide free (AB238948)

USP22 was immunoprecipitated from 0.35 mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab195289 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab195289 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1 : HeLa whole cell lysate, 10μg (Input).

Lane 2 : ab195289 IP in HeLa whole cell lysate.

Lane 3 : Rabbit IgG,monoclonal [EPR25A]- Isotype Control (ab172730) instead of ab195289 in HeLa whole cell lysate.

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

Exposure time : 3 seconds.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab195289).

All lanes:

Immunoprecipitation - Anti-USP22 antibody [EPR18945] (<a href='/en-us/products/primary-antibodies/usp22-antibody-epr18945-ab195289'>ab195289</a>)

Predicted band size: 60 kDa

false

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-USP22 antibody [EPR18945] - BSA and Azide free (AB238948)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling USP22 with ab195289 at 1/1000 followed by a Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) Ready to use. Nuclear staining on mouse cerebrum. The section was incubated with ab195289 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) Ready to use.

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab195289).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-USP22 antibody [EPR18945] - BSA and Azide free (AB238948)

Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling USP22 with ab195289 at 1/1000 followed by a Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) Ready to use. Nuclear staining on rat cerebrum. The section was incubated with ab195289 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) Ready to use.

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab195289).

• WB

Lab

Western blot - Anti-USP22 antibody [EPR18945] - BSA and Azide free (AB238948)

False colour image of Western blot : Anti-USP22 antibody [EPR18945] staining at 1/2000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab195289 was shown to bind specifically to USP22. A band was observed at 59 kDa in wild-type HeLa cell lysates with no signal observed at this size in usp22 knockout cell line ab264888 (knockout cell lysate ab257789). To generate this image, wild-type and usp22 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 fluorescent western blot (TBS-based) blocking solution 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-USP22 antibody [EPR18945] (<a href='/en-us/products/primary-antibodies/usp22-antibody-epr18945-ab195289'>ab195289</a>) at 1/2000 dilution

Lane 1:

Western blot - Human USP22 knockout HeLa cell lysate (<a href='/en-us/products/cell-lysates/human-usp22-knockout-hela-cell-lysate-ab257789'>ab257789</a>)

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

USP22 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human USP22 knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-usp22-knockout-hela-cell-line-ab264888'>ab264888</a>)

Predicted band size: 60 kDa

Observed band size: 59 kDa

false

• WB

Supplier Data

Western blot - Anti-USP22 antibody [EPR18945] - BSA and Azide free (AB238948)

Lane 1 : Wild-type HAP1 whole cell lysate (20 μg)
Lane 2 : USP22 knockout HAP1 whole cell lysate (20 μg)
Lane 3 : HEK293 whole cell lysate (20 μg)
Lane 4 : HeLa whole cell lysate (20 μg)

Lanes 1 - 4 : Merged signal (red and green). Green - ab195289 observed at 60 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab195289 was shown to specifically react with USP22 in wild-type HAP1 cells. No band was observed when knockout samples were examined. Wild-type and USP22 knockout samples were subjected to SDS-PAGE. ab195289 and ab9484 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at a 1/2000 dilution and 1/20,000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) secondary antibodies at 1/20,000 dilution for 1 hour at room temperature before imaging.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab195289).

All lanes:

Western blot - Anti-USP22 antibody [EPR18945] - BSA and Azide free (ab238948)

Predicted band size: 60 kDa

false