Overview

  • Product name

    Anti-RPA70 antibody [EPR3472] - BSA and Azide free
    See all RPA70 primary antibodies
  • Description

    Rabbit monoclonal [EPR3472] to RPA70 - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

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

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human RPA70 aa 1-100. The exact sequence is proprietary.

  • General notes

    ab239890 is the carrier-free version of ab79398 This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    Ab239890 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

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

    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 ab239890 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.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

RIP Use at an assay dependent concentration. PubMed: 21957289
WB Use at an assay dependent concentration. Detects a band of approximately 70 kDa (predicted molecular weight: 70 kDa).
IP Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration. 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.
  • Information by UniProt
  • Database links

  • 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

  • Immunocytochemistry/Immunofluorescence analysis of A549 cells labelling 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).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab79398).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cervix carcinoma tissue labelling 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab79398).

  • Flow Cytometry analysis of HeLa cells labelling 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 - Unlabelled control, cells without incubation with primary and secondary antibodies.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab79398).

  • ab79398 (purified) at 1/20 immunoprecipitating RPA70 in HeLa whole cell lysate. 

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

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

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

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

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab79398).

  • Unpurified ab79398 staining RPA70 in U2OS 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab79398).

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

  • Overlay histogram showing HeLa cells stained with unpurified ab79398 (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 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab79398).

References

ab239890 has not yet been referenced specifically in any publications.

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