Anti-RPA32/RPA2 (phospho T21) 抗体 [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
Related conjugates and formulations
製品の概要
-
製品名
Anti-RPA32/RPA2 (phospho T21) antibody [EPR2846(2)]
RPA32/RPA2 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EPR2846(2)] to RPA32/RPA2 (phospho T21) -
由来種
Rabbit -
特異性
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.
-
アプリケーション
適用あり: Dot blot, WB, ELISAmore details
適用なし: Flow Cyt,ICC/IF or IHC-P -
種交差性
交差種: Mouse, Rat, Human -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
-
ポジティブ・コントロール
- WB: HeLa whole cell lysate treated with Calyculin A or Camptothecin. Dot Blot: RPA32/RPA2 (pT21) phospho peptide.
-
特記事項
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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
製品の特性
-
製品の状態
Liquid -
保存方法
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C. -
バッファー
pH: 7.20
Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant -
Concentration information loading...
-
精製度
Protein A purified -
ポリ/モノ
モノクローナル -
クローン名
EPR2846(2) -
アイソタイプ
IgG -
研究分野
関連製品
-
Alternative Versions
-
Compatible Secondaries
-
Isotype control
-
Recombinant Protein
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab109394の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
アプリケーション | Abreviews | 特記事項 |
---|---|---|
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.
|
特記事項 |
---|
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. |
ターゲット情報
-
機能
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. -
翻訳後修飾
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. -
細胞内局在
Nucleus. Nucleus > PML body. Also present in PML nuclear bodies. Redistributes to discrete nuclear foci upon DNA damage. - Information by UniProt
-
参照データベース
- Entrez Gene: 6118 Human
- Entrez Gene: 19891 Mouse
- Entrez Gene: 59102 Rat
- Omim: 179836 Human
- SwissProt: P15927 Human
- SwissProt: Q62193 Mouse
- SwissProt: Q63528 Rat
- Unigene: 703070 Human
see all -
別名
- 60S acidic ribosomal protein P1 antibody
- AA409079 antibody
- AI325195 antibody
see all
画像
-
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 secondsBlocking/Diluting buffer and concentration: 5% NFDM/TBST
-
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 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.
-
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.
-
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.
-
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 secondsBlocking 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?
データシートおよび資料
-
SDS download
-
Datasheet download
参考文献 (26)
ab109394 は 26 報の論文で使用されています。
- 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
- Llorens-Agost M et al. POLθ-mediated end joining is restricted by RAD52 and BRCA2 until the onset of mitosis. Nat Cell Biol 23:1095-1104 (2021). PubMed: 34616022
- Wu X et al. Enhancer of mRNA Decapping protein 4 (EDC4) interacts with replication protein a (RPA) and contributes to Cisplatin resistance in cervical Cancer by alleviating DNA damage. Hereditas 157:41 (2020). PubMed: 33054858