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重组RabMAb

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

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

Conjugates logo Related conjugates and formulations

Carrier Free

概述

  • 产品名称

    Anti-RPA32/RPA2 (phospho T21)抗体[EPR2846(2)]
    参阅全部 RPA32/RPA2 一抗

  • 描述

    兔单克隆抗体[EPR2846(2)] to RPA32/RPA2 (phospho T21)

  • 宿主

    Rabbit

  • 特异性

    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.

  • 经测试应用

    适用于: Dot blot, WB, ELISAmore details
    不适用于: Flow Cyt,ICC/IF or IHC-P

  • 种属反应性

    与反应: Mouse, Rat, Human

  • 免疫原

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

  • 阳性对照

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

  • 常规说明

    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.

性能

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab109394于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

应用 Ab评论 说明
Dot blot
1/1000.
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.
ELISA
Use at an assay dependent concentration.
说明
Dot blot
1/1000.
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.
ELISA
Use at an assay dependent concentration.
应用说明
Is unsuitable for Flow Cyt,ICC/IF or IHC-P.

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

    see all

  • 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

  • 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)

文献 (28)

发表研究结果有使用 ab109394？请让我们知道，以便我们可以引用本数据表中的参考文章。

ab109394 被引用在 28 文献中.

  • Menolfi D  et al. ATR kinase supports normal proliferation in the early S phase by preventing replication resource exhaustion. Nat Commun 14:3618 (2023). PubMed: 37336885
  • Moore CE  et al. RFWD3 promotes ZRANB3 recruitment to regulate the remodeling of stalled replication forks. J Cell Biol 222:N/A (2023). PubMed: 37036693
  • Elbakry A  et al. ATRX and RECQ5 define distinct homologous recombination subpathways. Proc Natl Acad Sci U S A 118:N/A (2021). PubMed: 33431668
  • Mohr L  et al. ER-directed TREX1 limits cGAS activation at micronuclei. Mol Cell 81:724-738.e9 (2021). PubMed: 33476576
  • Shi J  et al. MTA2 sensitizes gastric cancer cells to PARP inhibition by induction of DNA replication stress. Transl Oncol 14:101167 (2021). PubMed: 34280886
View all Publications for this product

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Immunocytochemistry/ Immunofluorescence abreview for Anti-RPA32/RPA2 (phospho T21) antibody [EPR2846(2)]

 Good
Abreviews
This product is known to not work in this application or species.
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (U2OS)
Permeabilization
Yes - Triton 0.3%
Specification
U2OS
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Fixative
Paraformaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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提交于 Jan 24 2013

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