Astana Biomed Group, an authorized Abcam distributor in Central Asia

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

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR2846(2)] to RPA32/RPA2 (phospho T21)
  • Suitable for: Dot blot, WB, ELISA
  • Reacts with: Mouse, Rat, Human, Recombinant fragment

Overview

  • Product name

    Anti-RPA32/RPA2 (phospho T21) antibody [EPR2846(2)]
    See all RPA32/RPA2 primary antibodies

  • Description

    Rabbit monoclonal [EPR2846(2)] to RPA32/RPA2 (phospho T21)

  • Host species

    Rabbit

  • Specificity

    ab109394 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.

    This antibody cross-reactivates with non-phospho substrate in specific situations.

  • Tested Applications & Species

    Application Species
    Dot
    Human
    ELISA
    Recombinant fragment
    WB
    Mouse
    Rat
    Human
    See all applications and species data

  • Immunogen

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

  • Positive control

    • WB: HeLa whole cell lysate treated with Calyculin A or Camptothecin. Dot Blot: RPA32/RPA2 (pT21) phospho peptide.

  • 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.

Properties

Images

  • Western blot - Anti-RPA32/RPA2 (phospho T21) antibody [EPR2846(2)] (ab109394)
    Western blot - Anti-RPA32/RPA2 (phospho T21) antibody [EPR2846(2)] (ab109394)
    All lanes : Anti-RPA32/RPA2 (phospho T21) antibody [EPR2846(2)] (ab109394) at 1/5000 dilution

    Lane 1 : MDA-MB-231 (Human breast adenocarcinoma epithelial cell) whole cell lysates
    Lane 2 : MDA-MB-231 (Human breast adenocarcinoma epithelial cell) whole cell lysates. Then the membrane was incubated with phosphatase.

    Lysates/proteins at 15 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 29 kDa
    Observed band size: 32,36 kDa
    why is the actual band size different from the predicted?


    Exposure time: 30 seconds


    Blocking/Diluting buffer and concentration: 5% NFDM/TBST

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

    Direct ELISA antigen dose-response curve using purified ab109394.

    Primary antibody: ab109394 (0-1000nweg/ml).

    Antigens: P1: Human RPA32/RPA2 (phospho T21) at 10ng/ml and 1ng/ml; NP: non-phospho at 10ng/ml and 1 ng/ml.

    Secondary antibody: Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) at 1/2500 dilution.

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

    Dot blot analysis of human c-Myb RPA32/RPA2 (pT21) phospho peptide (Lane 1) and RPA32/RPA2 non-phospho peptide (Lane 2) labeling RPA32/RPA2 (phospho T21) with purified ab109394 at a dilution of 1/5000. ab97051 (Peroxidase conjugated goat anti-rabbit IgG (H+L)) was used as the secondary antibody at a dilution of 1/2500.

    Blocking and dilution buffer: 5% NFDM/TBST.

    Exposure time: 3 minutes.

  • Western blot - Anti-RPA32/RPA2 (phospho T21) antibody [EPR2846(2)] (ab109394)
    Western blot - Anti-RPA32/RPA2 (phospho T21) antibody [EPR2846(2)] (ab109394)
    All lanes : Anti-RPA32/RPA2 (phospho T21) antibody [EPR2846(2)] (ab109394) at 1/2000 dilution

    Lane 1 : C6 (Rat glial tumor glial cell) whole cell lysates
    Lane 2 : C6 (Rat glial tumor glial cell) treated with 100nM calyculin A for 60 minutes whole cell lysates
    Lane 3 : C6 (Rat glial tumor glial cell) treated with 100nM calyculin A for 60 minutes whole cell lysates.Then the membrane was incubated with phosphatase.

    Lysates/proteins at 15 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 29 kDa
    Observed band size: 32,36 kDa why is the actual band size different from the predicted?



    Blocking/Diluting buffer and concentration:5% NFDM/TBST.

    Exposure time: 110 seconds.

  • Western blot - Anti-RPA32/RPA2 (phospho T21) antibody [EPR2846(2)] (ab109394)
    Western blot - Anti-RPA32/RPA2 (phospho T21) antibody [EPR2846(2)] (ab109394)
    All lanes : Anti-RPA32/RPA2 (phospho T21) antibody [EPR2846(2)] (ab109394) at 1/2000 dilution

    Lane 1 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates
    Lane 2 : NIH/3T3 (Mouse embryonic fibroblast) treated with 100ng/ml calyculin A for 30 minutes whole cell lysates
    Lane 3 : NIH/3T3 (Mouse embryonic fibroblast) treated with 100ng/ml calyculin A for 60 3minutes whole cell lysates.Then the membrane was incubated with phosphatase

    Lysates/proteins at 15 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 29 kDa
    Observed band size: 32,36 kDa why is the actual band size different from the predicted?



    Blocking/Diluting buffer and concentration: 5% NFDM/TBST.

    Exposure time: 180 seconds.

  • Western blot - Anti-RPA32/RPA2 (phospho T21) antibody [EPR2846(2)] (ab109394)
    Western blot - Anti-RPA32/RPA2 (phospho T21) antibody [EPR2846(2)] (ab109394)
    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
    All lanes : 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 - Anti-RPA32/RPA2 (phospho T21) antibody [EPR2846(2)] (ab109394)
    Dot Blot - Anti-RPA32/RPA2 (phospho T21) antibody [EPR2846(2)] (ab109394)

    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.

  • Western blot - Anti-RPA32/RPA2 (phospho T21) antibody [EPR2846(2)] (ab109394)
    Western blot - Anti-RPA32/RPA2 (phospho T21) antibody [EPR2846(2)] (ab109394)
    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?

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

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