Recombinant
RabMAb

Recombinant Anti-Pirh2 antibody [EPR18553] (ab189907)

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Overview

  • Product name

    Anti-Pirh2 antibody [EPR18553]
    See all Pirh2 primary antibodies

  • Description

    Rabbit monoclonal [EPR18553] to Pirh2

  • Host species

    Rabbit

  • Tested applications

    Suitable for: ICC/IF, IP, WBmore details

  • Species reactivity

    Reacts with: Mouse, Human

  • Immunogen

    Synthetic peptide within Human Pirh2 aa 150-250. The exact sequence is proprietary.
    Database link: Q96PM5

  • Positive control

    • WB: HeLa, HepG2, Daudi, HEK-293, HCT 116, LNCaP, RAW 264.7 and NIH/3T3 whole cell lysates; Human fetal heart, fetal kidney and fetal spleen lysates; Mouse kidney and spleen lysates. ICC/IF: HeLa and HEK-293 cells. IP: HeLa whole cell lysate.

  • 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

Our Abpromise guarantee covers the use of ab189907 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
ICC/IF 1/500.
IP 1/50.
WB 1/1000. Detects a band of approximately 30 kDa (predicted molecular weight: 30 kDa).

Target

  • Function

    Mediates E3-dependent ubiquitination and proteasomal degradation of target proteins, including p53/TP53, HDAC1 and CDKN1B. Preferentially acts on tetrameric p53/TP53. Contributes to the regulation of CDKN1B and p53/TP53 levels, and thereby contributes to the regulation of the cell cycle progression. Increases AR transcription factor activity.

  • Pathway

    Protein modification; protein ubiquitination.

  • Sequence similarities

    Contains 1 CHY-type zinc finger.
    Contains 1 CTCHY-type zinc finger.
    Contains 1 RING-type zinc finger.

  • Post-translational
    modifications

    Subject to ubiquitination and proteasomal degradation. Interaction with PLAGL2 or KAT5 enhances protein stability.

  • Cellular localization

    Nucleus. Nucleus speckle. Cytoplasm.
  • Information by UniProt

  • Database links

    • Alternative names

      • Androgen receptor N terminal interacting protein antibody
      • Androgen receptor N-terminal-interacting protein antibody
      • ARNIP antibody
      • CH-rich-interacting match with PLAG1 antibody
      • CHIMP antibody
      • E3 ubiquitin-protein ligase Pirh2 antibody
      • hARNIP antibody
      • hPirh2 antibody
      • p53 induced protein with a RING H2 domain antibody
      • p53-induced RING-H2 protein antibody
      • PIRH2E antibody
      • PIRH2F antibody
      • PRO1996 antibody
      • RCHY1 antibody
      • Ring finger and CHY zinc finger domain containing 1 E3 ubiquitin protein ligase antibody
      • RING finger and CHY zinc finger domain-containing protein 1 antibody
      • RING finger protein 199 antibody
      • RNF199 antibody
      • ZCHY antibody
      • ZFP 363 antibody
      • zinc finger CHY type antibody
      • Zinc finger protein 363 antibody
      • ZN363_HUMAN antibody
      • ZNF363 antibody
      see all

    Images

    • Western blot - Anti-Pirh2 antibody [EPR18553] (ab189907)
      Western blot - Anti-Pirh2 antibody [EPR18553] (ab189907)
      All lanes : Anti-Pirh2 antibody [EPR18553] (ab189907) at 1/1000 dilution

      Lane 1 : Wild-type HeLa cell lysate
      Lane 2 : RCHY1 knockout HeLa cell lysate
      Lane 3 : HepG2 cell lysate
      Lane 4 : Daudi cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 30 kDa



      Lanes 1- 4: Merged signal (red and green). Green - ab189907 observed at 30 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

      ab189907 was shown to react with RCHY1 in wild-type HeLa cells in Western blot. Loss of signal was observed when knockout sample ab258171 was used. Wild-type HeLa and RCHY1 knockout HeLa whole cell lysates were subjected to SDS-PAGE. ab189907 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

    • Immunocytochemistry/ Immunofluorescence - Anti-Pirh2 antibody [EPR18553] (ab189907)
      Immunocytochemistry/ Immunofluorescence - Anti-Pirh2 antibody [EPR18553] (ab189907)

      Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling Pirh2 with ab189907 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

      Confocal image showing nuclear and cytoplasmic staining on HeLa cell line.

      The nuclear counterstain is DAPI (blue).

      Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).

      The negative controls are as follows:-

      -ve control 1: ab189907 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.

      -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

    • Immunocytochemistry/ Immunofluorescence - Anti-Pirh2 antibody [EPR18553] (ab189907)
      Immunocytochemistry/ Immunofluorescence - Anti-Pirh2 antibody [EPR18553] (ab189907)

      Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HEK-293 (Human epithelial cells from embryonic kidney) cells labeling Pirh2 with ab189907 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

      Confocal image showing nuclear and cytoplasmic staining on HEK-293 cell line.

      The nuclear counterstain is DAPI (blue).

      Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).

      The negative controls are as follows:-

      -ve control 1: ab189907 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.

      -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

    • Western blot - Anti-Pirh2 antibody [EPR18553] (ab189907)
      Western blot - Anti-Pirh2 antibody [EPR18553] (ab189907)
      All lanes : Anti-Pirh2 antibody [EPR18553] (ab189907) at 1/1000 dilution

      Lane 1 : HEK-293 (Human epithelial cells from embryonic kidney) whole cell lysate
      Lane 2 : HCT 116 (Human colorectal carcinoma cell line) whole cell lysate
      Lane 3 : LNCaP (Human prostate cancer 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: 30 kDa
      Observed band size: 30 kDa


      Exposure time: 3 minutes


      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Western blot - Anti-Pirh2 antibody [EPR18553] (ab189907)
      Western blot - Anti-Pirh2 antibody [EPR18553] (ab189907)
      All lanes : Anti-Pirh2 antibody [EPR18553] (ab189907) at 1/1000 dilution

      Lanes 1 & 3 : Human fetal heart lysate
      Lane 2 : Human fetal kidney lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/10000 dilution

      Predicted band size: 30 kDa
      Observed band size: 30 kDa


      Exposure time: 30 seconds


      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Western blot - Anti-Pirh2 antibody [EPR18553] (ab189907)
      Western blot - Anti-Pirh2 antibody [EPR18553] (ab189907)
      All lanes : Anti-Pirh2 antibody [EPR18553] (ab189907) at 1/1000 dilution

      Lane 1 : Mouse kidney lysate
      Lane 2 : Mouse spleen 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: 30 kDa
      Observed band size: 30 kDa


      Exposure time: 3 minutes


      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Western blot - Anti-Pirh2 antibody [EPR18553] (ab189907)
      Western blot - Anti-Pirh2 antibody [EPR18553] (ab189907)
      All lanes : Anti-Pirh2 antibody [EPR18553] (ab189907) at 1/1000 dilution

      Lane 1 : RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate
      Lane 2 : NIH/3T3 (Mouse embyro fibroblast cells) 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: 30 kDa
      Observed band size: 30 kDa


      Exposure time: 3 minutes


      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Immunoprecipitation - Anti-Pirh2 antibody [EPR18553] (ab189907)
      Immunoprecipitation - Anti-Pirh2 antibody [EPR18553] (ab189907)

      Pirh2 was immunoprecipitated from 1mg of HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate with ab189907 at 1/50 dilution.

      Western blot was performed from the immunoprecipitate using ab189907 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 10ug (Input).

      Lane 2: ab189907 IP in HeLa whole cell lysate.

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab189907 in HeLa whole cell lysate.

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

      Exposure time: 30 seconds.

    References

    ab189907 has not yet been referenced specifically in any publications.

    Publishing research using ab189907? Please let us know so that we can cite the reference in this datasheet.

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