Anti-DNA polymerase eta抗体(ab17725)

概述

  • 产品名称Anti-DNA polymerase eta抗体
    参阅全部 DNA polymerase eta 一抗
  • 描述
    兔多克隆抗体to DNA polymerase eta
  • 特异性ab17725 specifically recognizes a band at ~80 kDa, the reported size of DNA polymerase eta, in MRC5 cells. This band is not seen in XP30RO cells, which lack endogenous DNA polymerase eta. Also, the band in MRC5 cells is no longer detected when the immunizing peptide is preincubated with the antibody before the membrane.
  • 经测试应用适用于: WB, ICC/IF, IHC-Pmore details
  • 种属反应性
    与反应: Human
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 650 to the C-terminus of Human DNA polymerase eta.

    (Peptide available as ab19077.)

  • 阳性对照
    • This antibody gave a positive signal in: (WB) HeLa: (IHC) Human Tonsil

性能

  • 形式Liquid
  • 存放说明Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • 存储溶液Preservative: 0.02% Sodium Azide
    Constituents: 1% BSA, PBS, pH 7.4
  • Concentration information loading...
  • 纯度Immunogen affinity purified
  • 克隆多克隆
  • 同种型IgG
  • 研究领域

相关产品

应用

Our Abpromise guarantee covers the use of ab17725 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
WB 1/400 - 1/2000. Detects a band of approximately 79-85 kDa (predicted molecular weight: 79 kDa).
ICC/IF Use at an assay dependent concentration. PubMed: 25421715
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

靶标

  • 功能DNA polymerase specifically involved in DNA repair. Plays an important role in translesion synthesis, where the normal high fidelity DNA polymerases cannot proceed and DNA synthesis stalls. Plays an important role in the repair of UV-induced pyrimidine dimers. Depending on the context, it inserts the correct base, but causes frequent base transitions and transversions. May play a role in hypermutation at immunoglobulin genes. Forms a Schiff base with 5'-deoxyribose phosphate at abasic sites, but does not have lyase activity. Targets POLI to replication foci.
  • 疾病相关Defects in POLH are the cause of xeroderma pigmentosum variant type (XPV) [MIM:278750]; also designated as XP-V. Xeroderma pigmentosum (XP) is an autosomal recessive disease due to deficient nucleotide excision repair. It is characterized by hypersensitivity of the skin to sunlight, followed by high incidence of skin cancer and frequent neurologic abnormalities. XPV shows normal nucleotide excision repair, but an exaggerated delay in recovery of replicative DNA synthesis. Most XPV patients do not develop clinical symptoms and skin neoplasias until a later age. Clinical manifestations are limited to photo-induced deterioration of the skin and eyes.
  • 序列相似性Belongs to the DNA polymerase type-Y family.
    Contains 1 umuC domain.
  • 结构域The catalytic core consists of fingers, palm and thumb subdomains, but the fingers and thumb subdomains are much smaller than in high-fidelity polymerases; residues from five sequence motifs of the Y-family cluster around an active site cleft that can accommodate DNA and nucleotide substrates with relaxed geometric constraints, with consequently higher rates of misincorporation and low processivity.
  • 细胞定位Nucleus. Accumulates at replication forks after DNA damage.
  • Information by UniProt
  • 数据库链接
  • 别名
    • DNA polymerase eta antibody
    • FLJ16395 antibody
    • FLJ21978 antibody
    • Polh antibody
    • POLH_HUMAN antibody
    • polymerase DNA directed eta antibody
    • RAD30 antibody
    • RAD30 homolog A antibody
    • RAD30A antibody
    • Xeroderma pigmentosum variant type protein antibody
    • XP V antibody
    • XPV antibody
    see all

Anti-DNA polymerase eta antibody 图像

  • Lanes 1, 2, 5 & 6 : Anti-DNA polymerase eta antibody (ab17725) at 1/400 dilution
    Lanes 3 - 4 : Anti-DNA polymerase eta antibody (ab17725) at 1/2000 dilution

    Lane 1 : XP30RO cell lysate (lacking endogenous DNA polymerase eta)
    Lane 2 : MRC5 cell lysate
    Lane 3 : XP30RO cell lysate (lacking endogenous DNA polymerase eta)
    Lane 4 : MRC5 cell lysate
    Lane 5 : XP30RO cell lysate(lacking endogenous DNA polymerase eta) with Human DNA polymerase eta peptide (ab19077) at 5 µl
    Lane 6 : MRC5 cell lysate with Human DNA polymerase eta peptide (ab19077) at 5 µl

    Secondary
    Goat anti-rabbit HRP at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 79 kDa
    Observed band size : 79 kDa


    Exposure time : 20 minutes

    60% confluent 6cm dish of each cell type was washed in PBS then scraped into 150µl of SDS-PAGE buffer, boiled for 5 minutes then sonicated for 10 minutes. 20µl of each sample was loaded in triplicate on a 6.5% polyacrylamide denaturing gel, then transferred to nitrocellulose. The membrane was blocked in 5% non-fat milk in PBS plus 0.1% Tween20 for 30 minutes. The primary antibody, ab17725, was incubated with the membrane in PBS plus 0.1% Tween20 and 5% milk overnight. Where the immunizing peptide was used, 5µl of the peptide was incubated with 5ul of antibody for ten minutes prior to dilution in 2ml of 5% milk in PBS plus 0.1% Tween20, which was then incubated with the blocked membrane overnight.

  • IHC image of DNA polymerase eta staining in human tonsil FFPE section, performed on a BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab17725, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
  • All lanes : Anti-DNA polymerase eta antibody (ab17725) at 2 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate with Human DNA polymerase eta peptide (ab19077) at 2 µg/ml

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Performed under reducing conditions.

    Predicted band size : 79 kDa
    Observed band size : 85 kDa (why is the actual band size different from the predicted?)


    Exposure time : 5 minutes
    Some batches of ab17725 observe a weaker 140 kDa band in addition to the target band at ~85 kDa. We are unsure as to the identify of the 140 kDa band and would welcome customer feedback.
  • All lanes : Anti-DNA polymerase eta antibody (ab17725) at 2 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate blocked with 5% milk
    Lane 2 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate blocked with 3% milk

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 79 kDa
    Observed band size : 85 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 140 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 20 minutes

Anti-DNA polymerase eta antibody (ab17725)参考文献

This product has been referenced in:
  • Sekimoto T  et al. Both high-fidelity replicative and low-fidelity Y-family polymerases are involved in DNA rereplication. Mol Cell Biol 35:699-715 (2015). Read more (PubMed: 25487575) »
  • Fattah FJ  et al. The transcription factor TFII-I promotes DNA translesion synthesis and genomic stability. PLoS Genet 10:e1004419 (2014). Human . Read more (PubMed: 24922507) »

See all 21 Publications for this product

Product Wall

Application Western blot
Loading amount 35 µg
Gel Running Conditions Reduced Denaturing (4-12% Bis-Tris)
Sample Mouse Cell lysate - whole cell (HEK293, HCT116, U2OS and Mouse Embryonic Fibroblas)
Specification HEK293, HCT116, U2OS and Mouse Embryonic Fibroblas
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Username

Abcam user community

Verified customer

提交于 Dec 23 2013

Application Western blot
Loading amount 35 µg
Gel Running Conditions Reduced Denaturing (4-12% Bis-Tris)
Sample Human Cell lysate - whole cell (HEK293, HCT116, U2OS and Mouse Embryonic Fibroblas)
Specification HEK293, HCT116, U2OS and Mouse Embryonic Fibroblas
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Username

Abcam user community

Verified customer

提交于 Dec 23 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Blocking step BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 2%
Sample Human Cell (U2OS)
Specification U2OS
Permeabilization Yes - 0.5% triton
Fixative Paraformaldehyde
Username

Dr. christophe lachaud

Verified customer

提交于 Jul 31 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Loading amount 25 µg
Gel Running Conditions Reduced Denaturing
Sample Human Cell lysate - whole cell (293 cells)
Specification 293 cells
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

Verified customer

提交于 Jul 24 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (U2OS cells)
Specification U2OS cells
Fixative Formaldehyde
Permeabilization Yes - np40
Blocking step BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: rt°C
Username

Abcam user community

Verified customer

提交于 Feb 15 2013

Thank you for your reply.

Indeed it looks like the antibody should have worked under these conditions. We have a different lot availible, and I am happy to provide this free of charge for your customer as they are covered by our Abpromise. Pl...

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Thank you for taking time to complete our questionnaire. I am sorry to hear that this antibody is not providing satisfactory results for your customer.



Having reviewed this case, I would appreciate if you can confirm some further...

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Ma collègue m'a transféré votre cas.

Concernant ab17725, nous n’avons que lot GR52626-1 en stock. Nous avons un nouveau lot en production qui devrait être disponible dans environ 2 mois s’il passe nos te...

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Thank you again for your patience with this.

We are currently transferringtwo vials from our lab stock to our distribution office, and they will be sent to you from there on the free of charge order # ***. I expect this shipment to arrive nex...

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