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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-USP22 antibody [EPR18945] (ab195289)

Reviews (1) Submit a question References (24)
Western blot - Anti-USP22 antibody [EPR18945] (ab195289)
  • Western blot - Anti-USP22 antibody [EPR18945] (ab195289)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-USP22 antibody [EPR18945] (ab195289)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-USP22 antibody [EPR18945] (ab195289)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-USP22 antibody [EPR18945] (ab195289)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-USP22 antibody [EPR18945] (ab195289)
  • Western blot - Anti-USP22 antibody [EPR18945] (ab195289)
  • Western blot - Anti-USP22 antibody [EPR18945] (ab195289)
  • Western blot - Anti-USP22 antibody [EPR18945] (ab195289)
  • Western blot - Anti-USP22 antibody [EPR18945] (ab195289)
  • Immunoprecipitation - Anti-USP22 antibody [EPR18945] (ab195289)
  • Anti-USP22 antibody [EPR18945] (ab195289)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR18945] to USP22
  • Suitable for: IHC-P, WB, IP
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 647 Carrier Free

Overview

  • Product name

    Anti-USP22 antibody [EPR18945]
    See all USP22 primary antibodies

  • Description

    Rabbit monoclonal [EPR18945] to USP22

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IHC-P, WB, IPmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Human fetal liver, fetal heart and fetal kidney lysates; HeLa, HEK-293, Jurkat, HepG2, MCF7, Neuro-2a, F9, C6, RAW 264.7, PC-12 and NIH/3T3 whole cell lysates; Mouse brain and spleen lysates; Rat brain and spleen lysates. IP: HeLa whole cell lysate. IHC-P: Human, mouse and rat cerebrum tissue; Human breast carcinoma tissue.

  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab195289 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IHC-P
1/1000.
WB
1/2000. Detects a band of approximately 60 kDa (predicted molecular weight: 60 kDa).
IP
1/40.
Notes
IHC-P
1/1000.
WB
1/2000. Detects a band of approximately 60 kDa (predicted molecular weight: 60 kDa).
IP
1/40.

Target

  • Function

    Histone deubiquitinating component of the transcription regulatory histone acetylation (HAT) complex SAGA. Catalyzes the deubiquitination of both histones H2A and H2B, thereby acting as a coactivator. Recruited to specific gene promoters by activators such as MYC, where it is required for transcription. Required for nuclear receptor-mediated transactivation and cell cycle progression.

  • Tissue specificity

    Moderately expressed in various tissues including heart and skeletal muscle, and weakly expressed in lung and liver.

  • Sequence similarities

    Belongs to the peptidase C19 family. UBP8 subfamily.
    Contains 1 UBP-type zinc finger.

  • Cellular localization

    Nucleus.
  • Information by UniProt

  • Database links

    • Alternative names

      • Deubiquitinating enzyme 22 antibody
      • KIAA1063 antibody
      • Ubiquitin carboxyl terminal hydrolase 22 antibody
      • Ubiquitin carboxyl-terminal hydrolase 22 antibody
      • Ubiquitin specific peptidase 22 antibody
      • Ubiquitin specific peptidase 3 like antibody
      • Ubiquitin specific processing protease 22 antibody
      • Ubiquitin specific protease 22 antibody
      • Ubiquitin thioesterase 22 antibody
      • Ubiquitin thiolesterase 22 antibody
      • Ubiquitin-specific-processing protease 22 antibody
      • UBP22_HUMAN antibody
      • USP 22 antibody
      • Usp22 antibody
      • USP3L antibody
      see all

    Images

    • Western blot - Anti-USP22 antibody [EPR18945] (ab195289)
      Western blot - Anti-USP22 antibody [EPR18945] (ab195289)
      All lanes : Anti-USP22 antibody [EPR18945] (ab195289) at 1/2000 dilution

      Lane 1 : Wild-type HeLa cell lysate
      Lane 2 : USP22 knockout HeLa cell lysate

      Performed under reducing conditions.

      Predicted band size: 60 kDa
      Observed band size: 59 kDa why is the actual band size different from the predicted?



      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.

    • Western blot - Anti-USP22 antibody [EPR18945] (ab195289)
      Western blot - Anti-USP22 antibody [EPR18945] (ab195289)

      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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-USP22 antibody [EPR18945] (ab195289)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-USP22 antibody [EPR18945] (ab195289)

      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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-USP22 antibody [EPR18945] (ab195289)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-USP22 antibody [EPR18945] (ab195289)

      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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-USP22 antibody [EPR18945] (ab195289)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-USP22 antibody [EPR18945] (ab195289)

      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) at Ready to use dilution. 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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-USP22 antibody [EPR18945] (ab195289)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-USP22 antibody [EPR18945] (ab195289)

      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. 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

    • Western blot - Anti-USP22 antibody [EPR18945] (ab195289)
      Western blot - Anti-USP22 antibody [EPR18945] (ab195289)

      Lane 1: Wild type HAP1 whole cell lysate (20 µg)
      Lane 2: USP22 knockout HAP1 cell lysate (20 μg)
      Lane 3: HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate (20 µg)

      ab195289 was shown to specifically recognize USP22  when USP22 knockout samples were used. Wild-type and knockout samples were subjected to SDS-PAGE. ab195289 (1/2000) and ab181602 (Rabbit anti GAPDH loading control) were incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100,000 dilution.  

    • Western blot - Anti-USP22 antibody [EPR18945] (ab195289)
      Western blot - Anti-USP22 antibody [EPR18945] (ab195289)
      All lanes : Anti-USP22 antibody [EPR18945] (ab195289) at 1/2000 dilution

      Lane 1 : Human fetal liver lysate
      Lane 2 : Human fetal heart lysate
      Lane 3 : Human fetal kidney lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

      Predicted band size: 60 kDa
      Observed band size: 60 kDa


      Exposure time: 3 minutes


      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Western blot - Anti-USP22 antibody [EPR18945] (ab195289)
      Western blot - Anti-USP22 antibody [EPR18945] (ab195289)
      All lanes : Anti-USP22 antibody [EPR18945] (ab195289) at 1/2000 dilution

      Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
      Lane 2 : HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
      Lane 3 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
      Lane 4 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate
      Lane 5 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate
      Lane 6 : Neuro-2a (Mouse neuroblastoma cell line) whole cell lysate
      Lane 7 : F9 (Mouse embryonic testicular cancer cell line) whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 60 kDa
      Observed band size: 60 kDa



      Blocking/Dilution buffer: 5% NFDM/TBST.

      Exposure times: Lane 1, 2, 3, 4, 5 and 6: 30 seconds; Lane 7: 15 seconds.

    • Western blot - Anti-USP22 antibody [EPR18945] (ab195289)
      Western blot - Anti-USP22 antibody [EPR18945] (ab195289)
      All lanes : Anti-USP22 antibody [EPR18945] (ab195289) at 1/2000 dilution

      Lane 1 : Mouse brain lysate
      Lane 2 : Mouse spleen lysate
      Lane 3 : Rat brain lysate
      Lane 4 : Rat spleen lysate
      Lane 5 : C6 (Rat glial tumor cell line) whole cell lysate
      Lane 6 : RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
      Lane 7 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
      Lane 8 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 60 kDa
      Observed band size: 60 kDa



      Blocking/Dilution buffer: 5% NFDM/TBST.

      Exposure times: Lane 1, 2, 3 and 4: 3 minutes; Lane 5, 6, 7 and 8: 30 seconds.

       

    • Immunoprecipitation - Anti-USP22 antibody [EPR18945] (ab195289)
      Immunoprecipitation - Anti-USP22 antibody [EPR18945] (ab195289)

      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.

    • Anti-USP22 antibody [EPR18945] (ab195289)
      Anti-USP22 antibody [EPR18945] (ab195289)

    References (24)

    Publishing research using ab195289? Please let us know so that we can cite the reference in this datasheet.

    ab195289 has been referenced in 24 publications.

    • Chang YS  et al. Upregulation of USP22 and ABCC1 during Sorafenib Treatment of Hepatocellular Carcinoma Contribute to Development of Resistance. Cells 11:N/A (2022). PubMed: 35203285
    • Yadav P  et al. M6A RNA Methylation Regulates Histone Ubiquitination to Support Cancer Growth and Progression. Cancer Res 82:1872-1889 (2022). PubMed: 35303054
    • Zhang S  et al. Targeting ferroptosis with miR-144-3p to attenuate pancreatic β cells dysfunction via regulating USP22/SIRT1 in type 2 diabetes. Diabetol Metab Syndr 14:89 (2022). PubMed: 35761309
    • Li Q  et al. PRDM1/BLIMP1 induces cancer immune evasion by modulating the USP22-SPI1-PD-L1 axis in hepatocellular carcinoma cells. Nat Commun 13:7677 (2022). PubMed: 36509766
    • Wei Y  et al. USP22 promotes melanoma and BRAF inhibitor resistance via YAP stabilization. Oncol Lett 21:394 (2021). PubMed: 33777217
    View all Publications for this product

    Customer reviews and Q&As

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    Immunocytochemistry/ Immunofluorescence abreview for Anti-USP22 antibody [EPR18945]

     Excellent
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HeLa)
    Permeabilization
    Yes - 0.5% Triton-X100 in PBS
    Specification
    HeLa
    Fixative
    Paraformaldehyde
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    DR. Kirk Mcmanus

    Verified customer

    Submitted Sep 30 2016

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