概述

  • 产品名称
    Anti-RPA32/RPA2抗体[9H8]
    参阅全部 RPA32/RPA2 一抗
  • 描述
    小鼠单克隆抗体[9H8] to RPA32/RPA2
  • 宿主
    Mouse
  • 经测试应用
    适用于: Flow Cyt, WB, IP, IHC-P, IHC-Fr, ICC/IFmore details
  • 种属反应性
    与反应: Human
  • 免疫原

    Full length protein (Human).

  • 阳性对照
    • WB: TE 671, SK N BE and HeLa whole cell lysates. IHC-P: Human tonsil tissue. Flow Cyt: HeLa cells.

性能

应用

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.

应用 Ab评论 说明
Flow Cyt Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
WB Use at an assay dependent concentration. Predicted molecular weight: 32 kDa.
IP Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration. 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).

靶标

  • 功能
    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.
  • 翻译后修饰
    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.
  • 细胞定位
    Nucleus. Nucleus > PML body. Also present in PML nuclear bodies. Redistributes to discrete nuclear foci upon DNA damage.
  • Information by UniProt
  • 数据库链接
    • 别名
      • 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

    图片

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RPA32/RPA2 antibody [9H8] (ab2175)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RPA32/RPA2 antibody [9H8] (ab2175)
      Immunohistochemical analysis of formalin-fixed, paraffin-embedded Human tonsil tissue, staining RPA32/RPA2 with ab2175. Staining was detected using DAB.
    • Immunocytochemistry/ Immunofluorescence - Anti-RPA32/RPA2 antibody [9H8] (ab2175)
      Immunocytochemistry/ Immunofluorescence - Anti-RPA32/RPA2 antibody [9H8] (ab2175)This image is courtesy of an Abreview submitted by Dr Kok-Lung Chan

      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

    • Western blot - Anti-RPA32/RPA2 antibody [9H8] (ab2175)
      Western blot - Anti-RPA32/RPA2 antibody [9H8] (ab2175)
      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
      All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

      Predicted band size: 32 kDa
      Observed band size: 32 kDa

    • Flow Cytometry - Anti-RPA32/RPA2 antibody [9H8] (ab2175)
      Flow Cytometry - Anti-RPA32/RPA2 antibody [9H8] (ab2175)This image is courtesy of an anonymous Abreview

      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

    • Flow Cytometry - Anti-RPA32/RPA2 antibody [9H8] (ab2175)
      Flow Cytometry - Anti-RPA32/RPA2 antibody [9H8] (ab2175)

      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.

    文献

    This product has been referenced in:
    • Haas KT  et al. Single-molecule localization microscopy reveals molecular transactions during RAD51 filament assembly at cellular DNA damage sites. Nucleic Acids Res 46:2398-2416 (2018). Read more (PubMed: 29309696) »
    • Pancholi NJ & Weitzman MD Serotype-specific restriction of wild-type adenoviruses by the cellular Mre11-Rad50-Nbs1 complex. Virology 518:221-231 (2018). Read more (PubMed: 29547809) »

    See all 90 Publications for this product

    发表研究结果有使用 ab2175？请让我们知道，以便我们可以引用本数据表中的参考文章。

    客户评价及客户问答

    提交评价 提交问题

    11 reviews or Q&A

    10
    Votes: Highest First

    Immunocytochemistry/ Immunofluorescence abreview for Anti-RPA32/RPA2 antibody [9H8]

     Good
    Abreviews
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HeLa)
    Permeabilization
    Yes - 0.5% triton X
    Specification
    HeLa
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 21°C
    Fixative
    Paraformaldehyde
    Read More
    abreview image
    Username

    Abcam user community

    Verified customer

    提交于 Feb 08 2017

    ChIP abreview for Anti-RPA32/RPA2 antibody [9H8]

     Good
    Abreviews
    Application
    ChIP
    Sample
    Human Cell lysate - whole cell (U2OS)
    Negative control
    cells without 4OHT treatment and no antibody for IP
    Specification
    U2OS
    Detection step
    Real-time PCR
    Type
    Cross-linking (X-ChIP)
    Duration of cross-linking step: 15 minute(s) and 0 second(s)
    Specification of the cross-linking agent: fromaldehyde
    Positive control
    4 hydroxy tamoxifen (4OHT) treated cells (double strand breaks formed)
    Read More
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    Username

    Dr. Federica Marini

    Verified customer

    提交于 Dec 02 2016

    Immunocytochemistry/ Immunofluorescence abreview for Anti-RPA32/RPA2 antibody [9H8]

     Good
    Abreviews
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (U2OS)
    Permeabilization
    Yes - 0.5% Triton X-100
    Specification
    U2OS
    Fixative
    Paraformaldehyde
    Read More
    abreview image
    Username

    Abcam user community

    Verified customer

    提交于 Oct 06 2016

    Western blot abreview for Anti-RPA32/RPA2 antibody [9H8]

     Good
    Abreviews
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (breast cancer cell line)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    10 µg
    Treatment
    HU 1mM 16hr
    Specification
    breast cancer cell line
    Blocking step
    Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More
    Username

    Abcam user community

    Verified customer

    提交于 Jul 11 2016

    Western blot abreview for Anti-RPA32/RPA2 antibody [9H8]

     Excellent
    Abreviews
    Application
    Western blot
    Loading amount
    20 µg
    Gel Running Conditions
    Reduced Denaturing (8%)
    Sample
    Human Cell lysate - whole cell (U2OS cells)
    Specification
    U2OS cells
    Treatment
    1 ´M CPT
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More
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    Username

    Dr. Remi Buisson

    Verified customer

    提交于 Dec 03 2014

    Immunoprecipitation abreview for Anti-RPA32/RPA2 antibody [9H8]

     Excellent
    Abreviews
    Application
    Immunoprecipitation
    Immuno-precipitation step
    Protein G
    Sample
    Human Cell lysate - nuclear (U2OS cells)
    Specification
    U2OS cells
    Total protein in input
    200 µg
    Read More
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    Username

    Dr. Remi Buisson

    Verified customer

    提交于 Nov 22 2014

    ChIP abreview for Anti-RPA32/RPA2 antibody [9H8]

     Excellent
    Abreviews
    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)
    Read More
    Username

    Abcam user community

    Verified customer

    提交于 Nov 27 2013

    Flow Cytometry abreview for Anti-RPA32/RPA2 antibody [9H8]

     Excellent
    Abreviews
    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
    Read More
    abreview image
    Username

    Abcam user community

    Verified customer

    提交于 Aug 17 2012

    Immunocytochemistry/ Immunofluorescence abreview for Anti-RPA32/RPA2 antibody [9H8]

     Good
    Abreviews
    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
    Read More
    abreview image
    Username

    Abcam user community

    Verified customer

    提交于 Oct 28 2011

    Immunocytochemistry/ Immunofluorescence abreview for Anti-RPA32/RPA2 antibody [9H8]

     Excellent
    Abreviews
    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
    Read More
    abreview image
    Username

    Dr. Kok-Lung Chan

    Verified customer

    提交于 Nov 08 2007

    1-10 of 11 Abreviews or Q&A

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

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