Overview

  • Product name

    Anti-RPA32/RPA2 antibody [EPR2877Y]
    See all RPA32/RPA2 primary antibodies
  • Description

    Rabbit monoclonal [EPR2877Y] to RPA32/RPA2
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Flow Cyt, WB, IP, IHC-P, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human RPA32/RPA2 (C terminal). The exact sequence is proprietary.

  • Positive control

    • IHC-P: Human breast carcinoma tissue, mouse and rat liver tissue WB: HeLa whole cell lysate (ab150035), HUVEC lysate, NIH3T3 cell lysate, C6 cell lysate ICC/IF: C6, HeLa, NIH/3T3 cells. FC: C6, HeLa, NIH/3T3 cells. IP: HeLa 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.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab76420 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
Flow Cyt Use at an assay dependent concentration.
WB 1/1000. Detects a band of approximately 29 kDa (predicted molecular weight: 29 kDa).

For unpurified format use at 1/5000 - 1/10000.

IP 1/20.

For unpurified format use at 1/40.

IHC-P 1/100.

See IHC antigen retrieval protocols.

ICC/IF 1/50.

Target

  • Function

    Required for DNA recombination, repair and replication. The activity of RP-A is mediated by single-stranded DNA binding and protein interactions.
    Functions as component of the alternative replication protein A complex (aRPA). aRPA binds single-stranded DNA and probably plays a role in DNA repair; it does not support chromosomal DNA replication and cell cycle progression through S-phase. In vitro, aRPA cannot promote efficient priming by DNA polymerase alpha but supports DNA polymerase delta synthesis in the presence of PCNA and replication factor C (RFC), the dual incision/excision reaction of nucleotide excision repair and RAD51-dependent strand exchange.
  • Post-translational
    modifications

    Phosphorylated in a cell-cycle-dependent manner (from the S phase until mitosis). Phosphorylated by ATR upon DNA damage, which promotes its translocation to nuclear foci. Can be phosphorylated in vitro by PRKDC/DNA-PK in the presence of Ku and DNA, and by CDK1.
  • Cellular localization

    Nucleus. Nucleus > PML body. Also present in PML nuclear bodies. Redistributes to discrete nuclear foci upon DNA damage.
  • Information by UniProt
  • Database links

  • Alternative names

    • 60S acidic ribosomal protein P1 antibody
    • AA409079 antibody
    • AI325195 antibody
    • AU020965 antibody
    • ik:tdsubc_2g1 antibody
    • M(2)21C antibody
    • MGC137236 antibody
    • OTTHUMP00000004008 antibody
    • p32 antibody
    • p34 antibody
    • RCJMB04_6d17 replication protein A2, 32kDa antibody
    • REPA2 antibody
    • Replication factor A protein 2 antibody
    • Replication protein A 32 kDa subunit antibody
    • Replication protein A 32kDa subunit antibody
    • Replication protein A 34 kDa subunit antibody
    • Replication protein A antibody
    • Replication Protein A2 (32kDa) antibody
    • Replication protein A2 antibody
    • Replication protein A2, 32kDa antibody
    • RF-A protein 2 antibody
    • Rf-A2 antibody
    • RFA antibody
    • RFA2_HUMAN antibody
    • RP-A p32 antibody
    • RP-A p34 antibody
    • RP21C antibody
    • RPA 2 antibody
    • RPA 32 antibody
    • RPA antibody
    • Rpa2 antibody
    • RPA32 antibody
    • RPA34 antibody
    • RpLP1 antibody
    • RpP2 antibody
    • xx:tdsubc_2g1 antibody
    • zgc:109822 antibody
    see all

Images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human breast cancer tissue sections labeling RPA32/RPA2 with purified ab76420 at 1/100 dilution (1 µg/ml). Heat mediated antigen retrieval was performed using heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

  • All lanes : Anti-RPA32/RPA2 antibody [EPR2877Y] (ab76420) at 1/1000 dilution (Purified)

    Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
    Lane 2 : C6 (Rat glial tumor glial cell) whole cell lysates
    Lane 3 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates

    Lysates/proteins at 15 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 29 kDa
    Observed band size: 29 kDa

  • Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling RPA32/RPA2 with purified ab76420 at 1/50 dilution (2.0 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1/200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

  • All lanes : Anti-RPA32/RPA2 antibody [EPR2877Y] (ab76420) at 1/10000 dilution (unpurified)

    Lane 1 : HeLa cell lysate
    Lane 2 : HUVEC lysate
    Lane 3 : NIH 3T3 cell lysate
    Lane 4 : C6 cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : HRP labelled goat anti-rabbit at 1/1000 dilution

    Predicted band size: 29 kDa
    Observed band size: 29 kDa

  • Flow Cytometry analysis of C6 (rat glial tumor glial cell) cells labeling RPA32/RPA2 with purified ab76420 at 1:200 dilution (0.58 µg/ml) - Red. Cells were fixed with 4% paraformaldehyde . A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1:2000 dilution. Isotype control - Rabbit monoclonal IgG (ab172730) - Black. Unlabeled control - Blue. Untreated cells - Green
  • ab76420 (purified) at 1/20 dilution (0.5ug) immunoprecipitating RPA32/RPA2 in HeLa whole cell lysate. HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10ug
    Lane 2 (+): ab76420 & HeLa whole cell lysate
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab76420 in HeLa whole cell lysate
    For western blotting, VeriBlot for IP secondary antibody (HRP) (ab131366) was at 1/1000 dilution.
    Blocking and diluting buffer: 5% NFDM/TBST.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Rat liver tissue sections labeling RPA32/RPA2 with purified ab76420 at 1/100 dilution (1 µg/ml). Heat mediated antigen retrieval was performed using heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Mouse liver tissue sections labeling RPA32/RPA2 with purified ab76420 at 1/100 dilution (1 µg/ml). Heat mediated antigen retrieval was performed using heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

  • Immunocytochemistry/ Immunofluorescence analysis of C6 (rat glial tumor glial cell) cells labeling RPA32/RPA2 with purified ab76420 at 1:50 (2.28 µg/ml). Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
  • Immunocytochemistry/ Immunofluorescence analysis of NIH/3T3 (mouse embryonic fibroblast) cells labeling RPA32/RPA2 with purified ab76420 at 1:50 (2.28 µg/ml). Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
  • Immunocytochemistry/ Immunofluorescence analysis of HeLa (human cervix adenocarcinoma epithelial cell) cells labeling RPA32/RPA2 with purified ab76420 at 1:50 (2.28 µg/ml). Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
  • Flow Cytometry analysis of NIH/3T3 (mouse embryonic fibroblast) cells labeling RPA32/RPA2 with purified ab76420 at 1:200 dilution (0.58 µg/ml) - Red. Cells were fixed with 4% paraformaldehyde . A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1:2000 dilution. Isotype control - Rabbit monoclonal IgG (ab172730) - Black. Unlabeled control - Blue. Untreated cells - Green
  • Flow Cytometry analysis of HeLa (human cervix adenocarcinoma epithelial cell) cells labeling RPA32/RPA2 with purified ab76420 at 1:200 dilution (0.58 µg/ml) - Red. Cells were fixed with 4% paraformaldehyde . A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1:2000 dilution. Isotype control - Rabbit monoclonal IgG (ab172730) - Black. Unlabeled control - Blue. Untreated cells - Green
  • Immunohistochemical analysis of paraffin-embedded human breast carcinoma using ab76420 (unpurified) at 1/250 dilution.

References

This product has been referenced in:

  • Liu H  et al. SCRE serves as a unique synaptonemal complex fastener and is essential for progression of meiosis prophase I in mice. Nucleic Acids Res N/A:N/A (2019). Read more (PubMed: 30949703) »
  • Bhattacharyya R  et al. Coralyne, a protoberberine alkaloid, causes robust photosenstization of cancer cells through ATR-p38 MAPK-BAX and JAK2-STAT1-BAX pathways. Chem Biol Interact 285:27-39 (2018). Read more (PubMed: 29486184) »
See all 7 Publications for this product

Customer reviews and Q&As

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (spermatocytes)
Permeabilization
No
Specification
spermatocytes
Blocking step
Serum as blocking agent for 30 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 27°C
Fixative
Paraformaldehyde

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Verified customer

Submitted Dec 15 2016

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