Anti-Ku70 抗体 [EPR4027] (ab92450)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR4027] to Ku70
- Suitable for: WB, IP, IHC-P, ICC/IF, Flow Cyt (Intra)
- Reacts with: Human
Related conjugates and formulations
製品の概要
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製品名
Anti-Ku70 antibody [EPR4027]
Ku70 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EPR4027] to Ku70 -
由来種
Rabbit -
アプリケーション
適用あり: WB, IP, IHC-P, ICC/IF, Flow Cyt (Intra)more details -
種交差性
交差種: Human -
免疫原
Synthetic peptide within Human Ku70 aa 500-600. The exact sequence is proprietary.
Database link: P12956 -
ポジティブ・コントロール
- WB: A549, 293T, A431, and HeLa lysates. IHC-P: human colon carcinoma, tonsil and testis tissue. ICC/IF: HeLa cells. Flow Cyt (intra): HeLa cells. IP: HeLa cells.
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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.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
製品の特性
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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: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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精製度
Protein A purified -
ポリ/モノ
モノクローナル -
クローン名
EPR4027 -
アイソタイプ
IgG -
研究分野
関連製品
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Alternative Versions
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Compatible Secondaries
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Isotype control
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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ab92450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 |
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WB | (1) |
1/1000 - 1/10000. Predicted molecular weight: 70 kDa.
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IP |
1/20.
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IHC-P |
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Antigen retrieval andthe use of an HRP/AP polymerized secondary antibody is recommended for enhanced staining. |
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ICC/IF |
1/100 - 1/250.
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Flow Cyt (Intra) |
1/10 - 1/100.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
特記事項 |
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WB
1/1000 - 1/10000. Predicted molecular weight: 70 kDa. |
IP
1/20. |
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. Antigen retrieval andthe use of an HRP/AP polymerized secondary antibody is recommended for enhanced staining. |
ICC/IF
1/100 - 1/250. |
Flow Cyt (Intra)
1/10 - 1/100. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
ターゲット情報
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機能
Single stranded DNA-dependent ATP-dependent helicase. Has a role in chromosome translocation. The DNA helicase II complex binds preferentially to fork-like ends of double-stranded DNA in a cell cycle-dependent manner. It works in the 3'-5' direction. Binding to DNA may be mediated by XRCC6. Involved in DNA non-homologous end joining (NHEJ) required for double-strand break repair and V(D)J recombination. The XRCC5/6 dimer acts as regulatory subunit of the DNA-dependent protein kinase complex DNA-PK by increasing the affinity of the catalytic subunit PRKDC to DNA by 100-fold. The XRCC5/6 dimer is probably involved in stabilizing broken DNA ends and bringing them together. The assembly of the DNA-PK complex to DNA ends is required for the NHEJ ligation step. Required for osteocalcin gene expression. Probably also acts as a 5'-deoxyribose-5-phosphate lyase (5'-dRP lyase), by catalyzing the beta-elimination of the 5' deoxyribose-5-phosphate at an abasic site near double-strand breaks. 5'-dRP lyase activity allows to 'clean' the termini of abasic sites, a class of nucleotide damage commonly associated with strand breaks, before such broken ends can be joined. The XRCC5/6 dimer together with APEX1 acts as a negative regulator of transcription. -
配列類似性
Belongs to the ku70 family.
Contains 1 Ku domain.
Contains 1 SAP domain. -
発生段階
Expression does not increase during promyelocyte differentiation. -
翻訳後修飾
Phosphorylation by PRKDC may enhance helicase activity. Phosphorylation of Ser-51 does not affect DNA repair. -
細胞内局在
Nucleus. Chromosome. - Information by UniProt
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参照データベース
- Entrez Gene: 2547 Human
- Omim: 152690 Human
- SwissProt: P12956 Human
- Unigene: 292493 Human
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別名
- 5''-deoxyribose-5-phosphate lyase Ku70 antibody
- 5''-dRP lyase Ku70 antibody
- 70 kDa subunit of Ku antigen antibody
see all
画像
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All lanes : Anti-Ku70 antibody [EPR4027] (ab92450) at 1/5000 dilution (Purified)
Lane 1 : A549 (Human lung carcinoma epithelial cell) whole cell lysates
Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
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: 70 kDa
Observed band size: 70 kDa -
Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Ku70 with purified ab92450 at 1/250 dilution (0.46 µg/ml). Cells were fixed in 100% Methanol and permeabilized with None. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon carcinoma tissue sections labeling Ku70 with purified ab92450 at 1/250 dilution (0.46 µg/ml). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 1 (pH 6.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody.
Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
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All lanes : Anti-Ku70 antibody [EPR4027] (ab92450) at 1/1000 dilution ((unpurified))
Lane 1 : HeLa cell lysate
Lane 2 : 293T cell lysate
Lane 3 : A549 cell lysate
Lane 4 : A431 cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution
Predicted band size: 70 kDa -
Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Ku70 with purified ab92450 at 1/20 dilution (5µg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluorr® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
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ab92450 (purified ) at 1/20 dilution (0.5ug) immunoprecipitating Ku70 in HeLa whole cell lysate.
Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10ug
Lane 2 (+): ab92450 & HeLa whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab92450 in HeLa whole cell lysateFor western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/1000 dilution.
Blocking and diluting buffer: 5% NFDM/TBST.
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Immunofluorescence staining of HeLa cells using unpurified ab92450 at 1/100 dilution.
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Overlay histogram showing HeLa cells stained with unpurified ab92450 (red line). The cells were fixed with 4% paraformaldehyde (10 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 (ab92450, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
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Paraffin embedded Human testis tissue (A) or Human tonsil tissue (B) were labelled with unpurified ab92450 at 1/100 dilution.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
プロトコール
データシートおよび資料
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SDS download
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Datasheet download
参考文献 (14)
ab92450 は 14 報の論文で使用されています。
- Balbo Pogliano C et al. The CDK1-TOPBP1-PLK1 axis regulates the Bloom's syndrome helicase BLM to suppress crossover recombination in somatic cells. Sci Adv 8:eabk0221 (2022). PubMed: 35119917
- Tirado-Class N et al. PHIP variants associated with Chung-Jansen syndrome disrupt replication fork stability and genome integrity. Cold Spring Harb Mol Case Stud 8:N/A (2022). PubMed: 35863899
- Gao C et al. Inactivating Mutations of the IK Gene Weaken Ku80/Ku70-Mediated DNA Repair and Sensitize Endometrial Cancer to Chemotherapy. Cancers (Basel) 13:N/A (2021). PubMed: 34065218
- Townsend A et al. DCAF14 promotes stalled fork stability to maintain genome integrity. Cell Rep 34:108669 (2021). PubMed: 33503431
- Burgess JT et al. Barrier-to-autointegration-factor (Banf1) modulates DNA double-strand break repair pathway choice via regulation of DNA-dependent kinase (DNA-PK) activity. Nucleic Acids Res 49:3294-3307 (2021). PubMed: 33660778