製品の概要

  • 製品名Anti-RPA70 antibody [EPR3472]
    RPA70 一次抗体 製品一覧
  • 製品の詳細
    Rabbit monoclonal [EPR3472] to RPA70
  • アプリケーション適用あり: ICC/IF, WB, IP, IHC-P, Flow Cyt, RIPmore details
  • 種交差性
    交差種: Human
  • 免疫原

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human RPA70 aa 1-100.

  • ポジティブ・コントロール
    • WB: A549, HEK293 and HeLa cell lysates IHC-P: Human cervical squamous cell carcinoma tissue. ICC/IF: A549 cells. Flow Cyt: HeLa cells. IP: HeLa whole cell lysate.
  • 特記事項

    This product is a recombinant rabbit monoclonal antibody.

    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.

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

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

     

    Alternative versions available:

    Anti-RPA70 antibody (Alexa Fluor® 488) [EPR3472] (ab199097)
    Anti-RPA70 antibody (Alexa Fluor® 647) [EPR3472] (ab199240)

製品の特性

アプリケーション

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.

アプリケーション Abreviews 特記事項
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 protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

Flow Cyt 1/80.

For unpurified use at 1/50.

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

RIP Use at an assay dependent concentration. PubMed: 21957289

ターゲット情報

  • 機能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.
  • 配列類似性Belongs to the replication factor A protein 1 family.
  • 翻訳後修飾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.
  • 細胞内局在Nucleus.
  • Information by UniProt
  • 参照データベース
  • 別名
    • 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

Anti-RPA70 antibody [EPR3472] 画像

  • Anti-RPA70 antibody [EPR3472] (ab79398) at 1/1000 dilution (purified) + A549 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 and dilution buffer: 5% NFDM/TBST.

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

    Lane 1 : HEK293 cell lysate
    Lane 2 : HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

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

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

    Blocking and dilution buffer: 5% NFDM/TBST.

  • 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.

  • 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).

  • 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.

  • 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.

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

    Lane 1 : A549 cell lysate
    Lane 2 : HeLa cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    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) analysis of human cervical squamous cell carcinoma labelling RPA70 with unpurified ab79398 at a dilution of 1/100.

  • 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.

    See Abreview

  • 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.

Anti-RPA70 antibody [EPR3472] (ab79398) 使用論文

This product has been referenced in:
  • Alabert C  et al. Nascent chromatin capture proteomics determines chromatin dynamics during DNA replication and identifies unknown fork components. Nat Cell Biol 16:281-93 (2014). Read more (PubMed: 24561620) »
  • Toledo LI  et al. ATR prohibits replication catastrophe by preventing global exhaustion of RPA. Cell 155:1088-103 (2013). WB, ICC/IF ; Human . Read more (PubMed: 24267891) »

See all 3 Publications for this product

Product Wall

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
Username

Dr. Remi Buisson

Verified customer

投稿 Nov 21 2014

Application Immunoprecipitation
Immuno-precipitation step Protein G
Sample Human Cell lysate - nuclear (Hela cells)
Specification Hela cells
Total protein in input 200 µg
Username

Dr. Remi Buisson

Verified customer

投稿 Nov 20 2014

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
Username

Abcam user community

Verified customer

投稿 Aug 26 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
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
Username

Abcam user community

Verified customer

投稿 Oct 03 2012

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