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: WB, IP, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Human USP22 aa 1-150. The exact sequence is proprietary.
    Database link: Q9UPT9

  • 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.
  • General notes

     

     

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

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

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
WB 1/2000. Detects a band of approximately 60 kDa (predicted molecular weight: 60 kDa).
IP 1/40.
ICC/IF Use at an assay dependent concentration.

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

  • 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.

  • Immunocytochemistry/ Immunofluorescence analysis of HeLa cells labeling USP22 with ab195289. Cells were fixed with paraformaldehyde and permeabilized with 0.5% Triton-X100 in PBS. Samples were incubated with primary antibody (1/500 in PBS) for 1 hour at 22°C. ab150081 a goat anti-rabbit IgG (H + L) Alexa Fluor® 488 was used as the secondary antibody at 1/200 dilution.

    See Abreview

  • 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.  

  • 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.

  • 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.

  • 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.

     

  • 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.

References

This product has been referenced in:

  • Zhang H  et al. USP22 promotes resistance to EGFR-TKIs by preventing ubiquitination-mediated EGFR degradation in EGFR-mutant lung adenocarcinoma. Cancer Lett 433:186-198 (2018). Read more (PubMed: 29981430) »
  • Gennaro VJ  et al. Control of CCND1 ubiquitylation by the catalytic SAGA subunit USP22 is essential for cell cycle progression through G1 in cancer cells. Proc Natl Acad Sci U S A 115:E9298-E9307 (2018). Read more (PubMed: 30224477) »
See all 2 Publications for this product

Customer reviews and Q&As

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Permeabilization
Yes - 0.5% Triton-X100 in PBS
Specification
HeLa
Fixative
Paraformaldehyde

Dr. Kirk Mcmanus

Verified customer

Submitted Sep 30 2016

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