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    rpa70-antibody-epr3472-ab79398.pdf

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Epigenetics and Nuclear Signaling DNA / RNA DNA Damage & Repair Nucl. Excision Repair
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RecombinantRabMAb

Recombinant Anti-RPA70 antibody [EPR3472] (ab79398)

  • Datasheet
  • SDS
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Western blot - Anti-RPA70 antibody [EPR3472] (ab79398)
  • Immunocytochemistry/ Immunofluorescence - Anti-RPA70 antibody [EPR3472] (ab79398)
  • Immunocytochemistry/ Immunofluorescence - Anti-RPA70 antibody [EPR3472] (ab79398)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RPA70 antibody [EPR3472] (ab79398)
  • Immunoprecipitation - Anti-RPA70 antibody [EPR3472] (ab79398)
  • Flow Cytometry (Intracellular) - Anti-RPA70 antibody [EPR3472] (ab79398)
  • Western blot - Anti-RPA70 antibody [EPR3472] (ab79398)
  • Western blot - Anti-RPA70 antibody [EPR3472] (ab79398)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RPA70 antibody [EPR3472] (ab79398)
  • Flow Cytometry (Intracellular) - Anti-RPA70 antibody [EPR3472] (ab79398)
  • Anti-RPA70 antibody [EPR3472] (ab79398)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR3472] to RPA70
  • Suitable for: Flow Cyt (Intra), ICC/IF, WB, IP, IHC-P
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 647 APC Carrier Free HRP PE

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Overview

  • Product name

    Anti-RPA70 antibody [EPR3472]
    See all RPA70 primary antibodies
  • Description

    Rabbit monoclonal [EPR3472] to RPA70
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Flow Cyt (Intra), ICC/IF, WB, IP, IHC-Pmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: A549, HEK-293 and HeLa whole cell lysate (ab150035) IHC-P: Human cervical squamous cell carcinoma tissue. ICC/IF: A549 cells. Flow Cyt (intra): HeLa cells. IP: HeLa whole cell lysate (ab150035).
  • 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.

    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.

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR3472
  • Isotype

    IgG
  • Research areas

    • Epigenetics and Nuclear Signaling
    • DNA / RNA
    • DNA Damage & Repair
    • Nucl. Excision Repair
    • Epigenetics and Nuclear Signaling
    • DNA / RNA
    • DNA Damage & Repair
    • Homologous Recomb.
    • Epigenetics and Nuclear Signaling
    • DNA / RNA
    • DNA Synthesis
    • Other

Associated products

  • Alternative Versions

    • Alexa Fluor® 488 Anti-RPA70 antibody [EPR3472] (ab199097)
    • Alexa Fluor® 647 Anti-RPA70 antibody [EPR3472] (ab199240)
    • Anti-RPA70 antibody [EPR3472] - BSA and Azide free (ab239890)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96899)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • HeLa whole cell lysate (ab150035)
    • HeLa whole cell lysate (ab29545)
    • HEK-293 whole cell lysate (ab7902)
    • A549 whole cell lysate (ab7910)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab79398 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 (Intra)
1/80.

For unpurified use at 1/50.ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF (2)
1/100 - 1/250.
WB (1)
1/1000. Detects a band of approximately 70 kDa (predicted molecular weight: 70 kDa).

For unpurified use at 1/2000 - 1/5000.

IP (1)
1/10 - 1/20.
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

Notes
Flow Cyt (Intra)
1/80.

For unpurified use at 1/50.ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF
1/100 - 1/250.
WB
1/1000. Detects a band of approximately 70 kDa (predicted molecular weight: 70 kDa).

For unpurified use at 1/2000 - 1/5000.

IP
1/10 - 1/20.
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

Target

  • Function

    Plays an essential role in several cellular processes in DNA metabolism including replication, recombination and DNA repair. Binds and subsequently stabilizes single-stranded DNA intermediates and thus prevents complementary DNA from reannealing.
    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.
  • Sequence similarities

    Belongs to the replication factor A protein 1 family.
  • Post-translational
    modifications

    Phosphorylated upon DNA damage, probably by ATM or ATR.
    Sumoylated on lysine residues Lys-449 and Lys-577, with Lys-449 being the major site. Sumoylation promotes recruitment of RAD51 to the DNA damage foci to initiate DNA repair through homologous recombinaison. Desumoylated by SENP6.
  • Cellular localization

    Nucleus.
  • Target information above from: UniProt accession P27694 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 6117 Human
    • Omim: 179835 Human
    • SwissProt: P27694 Human
    • Unigene: 461925 Human
    • Alternative names

      • Dmrpa1 antibody
      • Drosophila Replication Protein A antibody
      • DRPA antibody
      • HSSB antibody
      • Human single stranded DNA binding protein antibody
      • MST075 antibody
      • MSTP075 antibody
      • p70 antibody
      • REPA1 antibody
      • Replication factor A antibody
      • Replication factor A protein 1 antibody
      • Replication protein A 70 kDa DNA-binding subunit antibody
      • Replication protein A 70kDa DNA binding subunit antibody
      • Replication protein A1 70kDa antibody
      • Replication protein A1 antibody
      • RF A antibody
      • RF-A protein 1 antibody
      • RFA antibody
      • RFA1_HUMAN antibody
      • RP A antibody
      • RP-A p70 antibody
      • RPA 70 antibody
      • RPA antibody
      • rpa1 antibody
      • Single stranded binding protein 70 antibody
      • Single-stranded DNA-binding protein antibody
      see all

    Images

    • Western blot - Anti-RPA70 antibody [EPR3472] (ab79398)
      Western blot - Anti-RPA70 antibody [EPR3472] (ab79398)
      All lanes : Anti-RPA70 antibody [EPR3472] (ab79398) at 1/4000 dilution (purified)

      Lane 1 : HEK-293 (Human epithelial cell line from embryonic kidney) cell lysate
      Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Peroxidase-conjugated goat anti-rabbit IgG, (H+L) at 1/1000 dilution

      Predicted band size: 70 kDa
      Observed band size: 70 kDa



      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Immunocytochemistry/ Immunofluorescence - Anti-RPA70 antibody [EPR3472] (ab79398)
      Immunocytochemistry/ Immunofluorescence - Anti-RPA70 antibody [EPR3472] (ab79398)This image is courtesy of an Abreview submitted by Remi Buisson

      Unpurified ab79398 staining RPA70 in U-2 OS (Human bone osteosarcoma epithelial cell line) cells by ICC/IF (Immunocytochemistry/immunofluorescence).

      Cells were fixed with paraformaldehyde, permeabilized with 0.5% Triton X-100 in PBS and blocked with 2% BSA for 1 hour at 25°C. Samples were incubated with primary antibody (1/500 in PBS + 0.5% Tween-20) for 2 hours at 25°C. A Cy3®-conjugated goat anti-rabbit IgG monoclonal (1/250) was used as the secondary antibody.

      See Abreview

    • Immunocytochemistry/ Immunofluorescence - Anti-RPA70 antibody [EPR3472] (ab79398)
      Immunocytochemistry/ Immunofluorescence - Anti-RPA70 antibody [EPR3472] (ab79398)

      Immunocytochemistry/Immunofluorescence analysis of A549 (Human lung carcinoma cell line) cells labeling RPA70 with purified ab79398 at 1/200.

      Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500) were also used.

      Control 1: primary antibody (1/200) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).

      Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RPA70 antibody [EPR3472] (ab79398)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RPA70 antibody [EPR3472] (ab79398)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cervix carcinoma tissue labeling RPA70 with purified ab79398 at 1/100.

      Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500).

      Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

    • Immunoprecipitation - Anti-RPA70 antibody [EPR3472] (ab79398)
      Immunoprecipitation - Anti-RPA70 antibody [EPR3472] (ab79398)

      ab79398 (purified) at 1/20 immunoprecipitating RPA70 in HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate. 

      Lane 1 (input): HeLa whole cell lysate (10 µg)

      Lane 2 (+): ab79398 + HeLa whole cell lysate (10 µg).

      Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab79398 in HeLa whole cell lysate.

      For western blotting, an HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG was used as the secondary antibody (1/1500).

      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Flow Cytometry (Intracellular) - Anti-RPA70 antibody [EPR3472] (ab79398)
      Flow Cytometry (Intracellular) - Anti-RPA70 antibody [EPR3472] (ab79398)

      Intracellular Flow Cytometry analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling RPA70 with purified ab79398 at 1/80 (red). 

      Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabeled control, cells without incubation with primary and secondary antibodies. 

       

       

    • Western blot - Anti-RPA70 antibody [EPR3472] (ab79398)
      Western blot - Anti-RPA70 antibody [EPR3472] (ab79398)
      Anti-RPA70 antibody [EPR3472] (ab79398) at 1/1000 dilution (purified) + A549 (Human lung carcinoma cell line) cell lysate at 20 µg

      Secondary
      Peroxidase-conjugated goat anti-rabbit IgG, (H+L) at 1/1000 dilution

      Predicted band size: 70 kDa
      Observed band size: 70 kDa



      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Western blot - Anti-RPA70 antibody [EPR3472] (ab79398)
      Western blot - Anti-RPA70 antibody [EPR3472] (ab79398)
      All lanes : Anti-RPA70 antibody [EPR3472] (ab79398) at 1/5000 dilution (unpurified)

      Lane 1 : A549 (Human lung carcinoma cell line) cell lysate
      Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution

      Predicted band size: 70 kDa
      Observed band size: 70 kDa

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RPA70 antibody [EPR3472] (ab79398)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RPA70 antibody [EPR3472] (ab79398)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cervical squamous cell carcinoma labelling RPA70 with unpurified ab79398 at a dilution of 1/100.

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    • Flow Cytometry (Intracellular) - Anti-RPA70 antibody [EPR3472] (ab79398)
      Flow Cytometry (Intracellular) - Anti-RPA70 antibody [EPR3472] (ab79398)

      Overlay histogram showing HeLa (Human epithelial cell line from cervix adenocarcinoma) cells stained with unpurifiedab79398 (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 (unpurified ab79398, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. 

      Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. 

      Acquisition of >5,000 events was performed. 

       

       

    • Anti-RPA70 antibody [EPR3472] (ab79398)
      Anti-RPA70 antibody [EPR3472] (ab79398)

    Protocols

    • Flow cytometry protocols
    • Immunoprecipitation protocols
    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (53)

    Publishing research using ab79398? Please let us know so that we can cite the reference in this datasheet.

    ab79398 has been referenced in 53 publications.

    • Herbert KJ  et al. Targeting TOPK sensitises tumour cells to radiation-induced damage by enhancing replication stress. Cell Death Differ 28:1333-1346 (2021). PubMed: 33168956
    • Zhang Y  et al. Pathways and signatures of mutagenesis at targeted DNA nicks. PLoS Genet 17:e1009329 (2021). PubMed: 33857147
    • Lezaja A  et al. RPA shields inherited DNA lesions for post-mitotic DNA synthesis. Nat Commun 12:3827 (2021). PubMed: 34158486
    • Ercilla A  et al. Physiological Tolerance to ssDNA Enables Strand Uncoupling during DNA Replication. Cell Rep 30:2416-2429.e7 (2020). PubMed: 32075739
    • Pedersen H  et al. Replication Protein A (RPA) Mediates Radio-Resistance of Glioblastoma Cancer Stem-Like Cells. Int J Mol Sci 21:N/A (2020). PubMed: 32111042
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-6 of 6 Abreviews or Q&A

    Immunocytochemistry/ Immunofluorescence abreview for Anti-RPA70 antibody [EPR3472]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (MCF10A cells)
    Permeabilization
    Yes - 0.2% Triton X-100 in PBS
    Specification
    MCF10A cells
    Blocking step
    10% FBS, 1%BSA, 0.1% Triton X-100 in PBS as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 11% · Temperature: 22°C
    Fixative
    Formaldehyde
    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.

    Abcam user community

    Verified customer

    Submitted Mar 17 2020

    ChIP abreview for Anti-RPA70 antibody [EPR3472]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    ChIP
    Sample
    Human Cell lysate - whole cell (HEK293E)
    Specification
    HEK293E
    Detection step
    Other
    Type
    Cross-linking (X-ChIP)
    Duration of cross-linking step: 15 minute(s) and 0 second(s)
    Specification of the cross-linking agent: formaldehyde
    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.

    Abcam user community

    Verified customer

    Submitted Dec 27 2019

    Immunocytochemistry/ Immunofluorescence abreview for Anti-RPA70 antibody [EPR3472]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C
    Sample
    Human Cell (U2OS)
    Specification
    U2OS
    Permeabilization
    Yes - 0.5% Triton /PBS
    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. Remi Buisson

    Verified customer

    Submitted Nov 21 2014

    Immunoprecipitation abreview for Anti-RPA70 antibody [EPR3472]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunoprecipitation
    Immuno-precipitation step
    Protein G
    Sample
    Human Cell lysate - nuclear (Hela cells)
    Specification
    Hela cells
    Total protein in input
    200 µg
    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. Remi Buisson

    Verified customer

    Submitted Nov 20 2014

    Flow Cytometry abreview for Anti-RPA70 antibody [EPR3472]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Flow Cytometry
    Fixation
    Paraformaldehyde
    Permeabilization
    Yes - 1xPBS + 0.2% Triton X-100
    Sample
    Human Cell (Human osteosarcoma U2OS cells)
    Specification
    Human osteosarcoma U2OS cells
    Gating Strategy
    Single cells discriminated by DAPI staining
    Preparation
    Cell harvesting/tissue preparation method: Trypsinization
    Sample buffer: 1xPBS + 1 mg/ml BSA + 1 microg/ml DAPI + 250 microg/ml RNase
    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.

    Abcam user community

    Verified customer

    Submitted Aug 26 2014

    Western blot abreview for Anti-RPA70 antibody [EPR3472]

    Good
    Abreviews
    Abreviews
    Application
    Western blot
    Sample
    Human Cell lysate - nuclear (Hela Cells)
    Loading amount
    50 µg
    Specification
    Hela Cells
    Gel Running Conditions
    Reduced Denaturing (10%)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
    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.

    Abcam user community

    Verified customer

    Submitted Oct 03 2012

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