Anti-RPA32/RPA2 (phospho T21) antibody [EPR2846(2)] (ab109394)

Overview

  • Product nameAnti-RPA32/RPA2 (phospho T21) antibody [EPR2846(2)]
    See all RPA32/RPA2 primary antibodies
  • Description
    Rabbit monoclonal [EPR2846(2)] to RPA32/RPA2 (phospho T21)
  • Specificityab109394 only detects RPA32/RPA2 phosphorylated at threonine 21. The antibody has been tested with milk (5%) and BSA (2%) as blocking reagents, but it provided better results with 5%milk.
  • Tested applicationsSuitable for: WB, IPmore details
    Unsuitable for: Flow Cyt,ICC/IF or IHC-P
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human RPA32/RPA2 (phospho T21). The exact sequence is proprietary.

  • Positive control
    • WB: HeLa cell lysates treated with camptothecin.
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

     

    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.

Properties

Applications

Our Abpromise guarantee covers the use of ab109394 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
WB 1/50000 - 1/200000. Detects a band of approximately 32 kDa (predicted molecular weight: 29 kDa).

The antibody has been tested with milk (5%) and BSA (2%) as blocking reagents, but it provided better results with 5%milk.

IP 1/10 - 1/100.
  • Application notesIs unsuitable for Flow Cyt,ICC/IF or IHC-P.
  • 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
      • 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

    Anti-RPA32/RPA2 (phospho T21) antibody [EPR2846(2)] images

    • All lanes : Anti-RPA32/RPA2 (phospho T21) antibody [EPR2846(2)] (ab109394) at 1/50000 dilution

      Lane 1 : HeLa whole cell lysate - untreated
      Lane 2 : HeLa whole cell lysate - treated with Calyculin A
      Lane 3 : HeLa whole cell lysate - treated with Calyculin A and Alkaline Phosphatase

      Lysates/proteins at 10 µg per lane.

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

      Predicted band size : 29 kDa
      Observed band size : 32 kDa (why is the actual band size different from the predicted?)


      Exposure time : 30 seconds

      Blocking and dilution buffer: 5% NFDM/TBST.

    • Dot blot analysis of RPA32/RPA2 (pT21) phospho peptide (lane 1) and RPA32/RPA2 non-phospho peptide (lane 2) labelling RPA32/RPA2 (phospho T21) with ab109394 at a dilution of 1/1000. A peroxidase-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/2500).

      Blocking and dilution buffer: 5% NFDM/TBST.

      Exposure time: 3 minutes.

    • All lanes : Anti-RPA32/RPA2 (phospho T21) antibody [EPR2846(2)] (ab109394) at 1/50000 dilution

      Lane 1 : HeLa cell lysates, untreated
      Lane 2 : HeLa cell lysates, treated with Camptothecin

      Lysates/proteins at 10 µg per lane.


      Predicted band size : 29 kDa
      Observed band size : 32 kDa (why is the actual band size different from the predicted?)

    References for Anti-RPA32/RPA2 (phospho T21) antibody [EPR2846(2)] (ab109394)

    This product has been referenced in:
    • Reczek CR  et al. The interaction between CtIP and BRCA1 is not essential for resection-mediated DNA repair or tumor suppression. J Cell Biol 201:693-707 (2013). Mouse . Read more (PubMed: 23712259) »
    • Kalan S  et al. LIM Protein Ajuba Participates in the Repression of the ATR-Mediated DNA Damage Response. Front Genet 4:95 (2013). Human . Read more (PubMed: 23755068) »

    See all 5 Publications for this product

    Product Wall

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Immunocytochemistry/ Immunofluorescence
    Sample Human Cell (U2OS)
    Specification U2OS
    Fixative Paraformaldehyde
    Permeabilization Yes - Triton 0.3%
    Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
    Username

    Abcam user community

    Verified customer

    Submitted Jan 24 2013

    Thank you for your email.

    This antibody has been guaranteed for Immunofluorescence i.e. ICC/IF. So the Abreview discount can not be provided. I can however provide you Image discount, when tested you need to submit an Abreview with ICC/IF imag...

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    Thank you for your reply.

    In the case of ab109394, we tested it on cells using a fluorophore conjugated secondary. So it is useful for your application.

    I would also like to remind you that all of our products are covered by our Abp...

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