Anti-DDB2抗体[2246C4a] (ab51017)

概述

  • 产品名称Anti-DDB2抗体[2246C4a]
    参阅全部 DDB2 一抗
  • 描述
    小鼠单克隆抗体[2246C4a] to DDB2
  • 经测试应用适用于: IHC-P, ICC/IF, WB, Dot blot, Flow Cytmore details
  • 种属反应性
    与反应: Human
  • 免疫原

    Recombinant fragment (N-terminal) Human.

  • 阳性对照
    • HeLa whole cell lysate

性能

应用

Our Abpromise guarantee covers the use of ab51017 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
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF Use a concentration of 10 µg/ml.
WB 1/50. Detects a band of approximately 45 kDa (predicted molecular weight: 48 kDa).
Dot blot Use at an assay dependent concentration.
Flow Cyt Use 1µg for 106 cells. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

靶标

  • 功能Required for DNA repair. Binds to DDB1 to form the UV-damaged DNA-binding protein complex (the UV-DDB complex). The UV-DDB complex may recognize UV-induced DNA damage and recruit proteins of the nucleotide excision repair pathway (the NER pathway) to initiate DNA repair. The UV-DDB complex preferentially binds to cyclobutane pyrimidine dimers (CPD), 6-4 photoproducts (6-4 PP), apurinic sites and short mismatches. Also appears to function as the substrate recognition module for the DCX (DDB1-CUL4-X-box) E3 ubiquitin-protein ligase complex DDB1-CUL4-ROC1 (also known as CUL4-DDB-ROC1 and CUL4-DDB-RBX1). The DDB1-CUL4-ROC1 complex may ubiquitinate histone H2A, histone H3 and histone H4 at sites of UV-induced DNA damage. The ubiquitination of histones may facilitate their removal from the nucleosome and promote subsequent DNA repair. The DDB1-CUL4-ROC1 complex also ubiquitinates XPC, which may enhance DNA-binding by XPC and promote NER. Isoform D1 and isoform D2 inhibit UV-damaged DNA repair.
  • 组织特异性Ubiquitously expressed; with highest levels in corneal endothelium and lowest levels in brain. Isoform D1 is highly expressed in brain and heart. Isoform D2, isoform D3 and isoform D4 are weakly expressed.
  • 通路Protein modification; protein ubiquitination.
  • 疾病相关Defects in DDB2 are a cause of xeroderma pigmentosum complementation group E (XP-E) [MIM:278740]; also known as xeroderma pigmentosum V (XP5). XP-E is a rare human autosomal recessive disease characterized by solar sensitivity, high predisposition for developing cancers on areas exposed to sunlight and, in some cases, neurological abnormalities.
  • 序列相似性Belongs to the WD repeat DDB2/WDR76 family.
    Contains 5 WD repeats.
  • 结构域The DWD box is required for interaction with DDB1.
  • 翻译后修饰Phosphorylation by ABL1 negatively regulate UV-DDB activity.
    Ubiquitinated by CUL4A in response to UV irradiation. Ubiquitination appears to both impair DNA-binding and promotes ubiquitin-dependent proteolysis. Degradation of DDB2 at sites of DNA damage may be a prerequisite for their recognition by XPC and subsequent repair. CUL4A-mediated degradation appears to be promoted by ABL1.
  • 细胞定位Nucleus. Accumulates at sites of DNA damage following UV irradiation.
  • Information by UniProt
  • 数据库链接
  • 别名
    • damage-specific DNA binding protein 2 antibody
    • Damage-specific DNA-binding protein 2 antibody
    • DDB p48 subunit antibody
    • Ddb2 antibody
    • DDB2_HUMAN antibody
    • DDBb antibody
    • DNA damage-binding protein 2 antibody
    • UV-damaged DNA-binding protein 2 antibody
    • UV-DDB 2 antibody
    • Xeroderma pigmentosum group E protei antibody
    see all

Anti-DDB2 antibody [2246C4a] 图像

  • Anti-DDB2 antibody [2246C4a] (ab51017) at 1/50 dilution + HeLa whole cell lysate at 50 µg

    Secondary
    Mouse IgG antibody at 1/2500 dilution
    Developed using the ECL technique

    Predicted band size : 48 kDa
    Observed band size : 45 kDa (why is the actual band size different from the predicted?)
  • IHC image of ab51017 staining in human normal cervical carcinoma formalin fixed paraffin embedded tissue section, performed on a Leica 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 ab51017, 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.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • ICC/IF image of ab51017 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab51017, 10µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • Overlay histogram showing HeLa cells stained with ab51017 (red line). The cells were fixed with 80% methanol (5 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. The cells were then incubated with the antibody (ab51017, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells ) used under the same conditions. Acquisition of >5,000 events was performed.

Anti-DDB2 antibody [2246C4a] (ab51017)参考文献

This product has been referenced in:
  • E X  et al. A novel DDB2-ATM feedback loop regulates human cytomegalovirus replication. J Virol 88:2279-90 (2014). Human . Read more (PubMed: 24335308) »
  • Luch A  et al. Low-dose formaldehyde delays DNA damage recognition and DNA excision repair in human cells. PLoS One 9:e94149 (2014). WB ; Human . Read more (PubMed: 24722772) »

See all 7 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (Fibroblasts)
Specification Fibroblasts
Fixative Paraformaldehyde
Permeabilization Yes - 0.5% Triton X-100
Blocking step Serum as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 20% · Temperature: 4°C
Username

Dr. Qien Wang

Verified customer

提交于 Jul 11 2012

Thank you very much for your interest in ab51017.

In general the testing discount program is meant for standard applications that have not been tested with a certain antibody yet. So ab51017 would not be eligible in IF staining since this is ...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - nuclear (GM00637 fibroblast cell line)
Loading amount 50 µg
Specification GM00637 fibroblast cell line
Gel Running Conditions Reduced Denaturing (10 %)
Blocking step Blocking buffer from another company as blocking agent for 30 minute(s) · Concentration: 5µg/mL · Temperature: 22°C
Username

Mrs. Annika Schäfer

Verified customer

提交于 Feb 10 2009

Thank you for your enquiry. I am sorry to hear that you are experiencing difficulties with this product ab51017 in western blot. Often it is possible to make suggestions that help resolve problems. We will happily offer technical support and in the ...

Read More

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"