Overview

  • Product nameAnti-RPA32/RPA2 antibody [9H8]See all RPA32/RPA2 primary antibodies ...
  • Description
    Mouse monoclonal [9H8] to RPA32/RPA2
  • SpecificityThis antibody reacts with a 32-34 kDa protein known as Replication Protein A (RPA).
  • Tested applicationsFlow Cyt, ChIP, WB, IP, IHC-P, IHC-Fr, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Full length protein (Human).

  • Positive control
    • Tonsil

Properties

Applications

Our Abpromise guarantee covers the use of ab2175 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 1µg for 106 cells.
ChIP Use at an assay dependent concentration.
WB Use at an assay dependent dilution. Predicted molecular weight: 32 kDa.
IP Use at an assay dependent dilution.
IHC-P Use at an assay dependent dilution.
IHC-Fr Use at an assay dependent dilution. This antibody may be diluted to a titer of 1/50-1/100 in an ABC method. We suggest an incubation period of 30 minutes at room temperature.
ICC/IF Use at an assay dependent concentration. Used at a dilution of 1/300 for 2 hrs on mouse MEF cells (see Abreview for further details).

Target

  • FunctionRequired 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 localizationNucleus. 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
    • HSSB antibody
    • ik:tdsubc_2g1 antibody
    • M(2)21C antibody
    • MGC137236 antibody
    • OTTHUMP00000004008 antibody
    • p32 antibody
    • p34 antibody
    • RCJMB04_6d17 replication protein A2, 32kDa antibody
    • REPA 2 antibody
    • REPA1 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 A1 (70kD) antibody
    • Replication Protein A2 (32kDa) antibody
    • Replication protein A2 32kD antibody
    • Replication protein A2 32kDa antibody
    • Replication protein A2 antibody
    • Replication protein A2, 32kDa antibody
    • RF A antibody
    • RF-A protein 2 antibody
    • Rf-A2 antibody
    • RFA antibody
    • RFA2_HUMAN antibody
    • RP A 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
    • RPA70 antibody
    • RpLP1 antibody
    • RpP2 antibody
    • xx:tdsubc_2g1 antibody
    • zgc:109822 antibody
    see all

Anti-RPA32/RPA2 antibody [9H8] images

  • ab2175 - immunohistochemistryFormalin fixed paraffin embedded human tonsil stained with RPA using ABC and AEC chromogen.
  • ab2175 at 1/200 staining human skin fibroblasts by ICC/IF. The cells were treated with 2mM hydroxyurea for 16 hours, formaldehyde fixed and permeabilized with 0.5% Triton X100. They were then incubated with the primary antibody for 1.5 hours at 37°C. A Cy3 ® conjugated sheep anti-mouse antibody was used as the secondary.

    See Abreview

  • All lanes : Anti-RPA32/RPA2 antibody [9H8] (ab2175) at 5 µg/ml

    Lane 1 : TE 671 (Human Rhabdomyosarcoma) Whole Cell Lysate
    Lane 2 : SK N BE (Human neuroblastoma) Whole Cell Lysate
    Lane 3 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Predicted band size : 32 kDa
    Observed band size : 32 kDa
  • Anti-RPA/32/RPA2 antibody [9H8] (ab2175) is used to measure DNA Double-Stranded Breaks resection in U2OS cells +/- Camptotechin (CPT).

    ab2175 is used at 1/200 dilution in 1xPBS + 0.2% Triton X-100 for 1 hour at 25°C.

    Secondary antibody: anti-mouse Molecular Probes Alexa Fluor® 488 Conjugated at 1/200 dilution.

    See Abreview

  • Immunohistochemical analysis of formalin-fixed, paraffin-embedded Human tonsil tissue, staining RPA32/RPA2 with ab2175. Staining was detected using DAB.
  • Overlay histogram showing HeLa cells stained with ab2175 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab2175, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

References for Anti-RPA32/RPA2 antibody [9H8] (ab2175)

This product has been referenced in:
  • Leung JW  et al. Nucleosome acidic patch promotes RNF168- and RING1B/BMI1-dependent H2AX and H2A ubiquitination and DNA damage signaling. PLoS Genet 10:e1004178 (2014). Human . Read more (PubMed: 24603765) »
  • Gao M  et al. Ago2 facilitates Rad51 recruitment and DNA double-strand break repair by homologous recombination. Cell Res 24:532-41 (2014). WB ; Human . Read more (PubMed: 24662483) »

See all 24 Publications for this product

Product Wall

Application ChIP
Detection step Real-time PCR
Sample Human Cell lysate - nuclear (hek293t)
Specification hek293t
Type Cross-linking (X-ChIP)
Duration of cross-linking step: 0 second(s)
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Abcam user community

Verified customer

Submitted Nov 27 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Flow Cytometry
Sample Human Cell (Human osteosarcoma U2OS cells)
Specification Human osteosarcoma U2OS cells
Preparation Cell harvesting/tissue preparation method: Trypsinization
Sample buffer: 1xPBS + 1 mg/ml BSA + 1 microg/ml DAPI + 250 microg/ml RNase
Fixation Paraformaldehyde
Permeabilization Yes - 1xPBS + 0.2% Triton X-100
Gating Strategy FS vs. SS for debris, DAPI area vs. linear for doublets
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Abcam user community

Verified customer

Submitted Aug 17 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HeLa)
Specification HeLa
Fixative Formaldehyde
Permeabilization Yes - 0.5% Triton X-100
Blocking step BSA as blocking agent for 15 minute(s) · Concentration: 5% · Temperature: 20°C
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Abcam user community

Verified customer

Submitted Oct 28 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (skin fibroblasts)
Specification skin fibroblasts
Fixative Formaldehyde
Permeabilization Yes - 0.5% Triton X100
Blocking step Serum as blocking agent for 20 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Dr. Kok-Lung Chan

Verified customer

Submitted Nov 08 2007

Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (MEF cells)
Specification MEF cells
Fixative Paraformaldehyde
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 5%
Username

Abcam user community

Verified customer

Submitted May 31 2006

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"