Recombinant Anti-DDB2 antibody [EPR9811] (ab181136)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR9811] to DDB2
- Suitable for: WB, IHC-P
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-DDB2 antibody [EPR9811]
See all DDB2 primary antibodies -
Description
Rabbit monoclonal [EPR9811] to DDB2 -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-Pmore details
Unsuitable for: Flow Cyt or ICC/IF -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HeLa, SH-SY5Y and Raji cell lysates; IHC-P: Human stomach carcinoma tissue.
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General notes
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.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR9811 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab181136 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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WB |
1/1000 - 1/10000. Detects a band of approximately 45 kDa (predicted molecular weight: 48 kDa).
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IHC-P |
1/2500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
See IHC antigen retrieval protocols. For unpurified use at 1/50 - 1/100. |
Notes |
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WB
1/1000 - 1/10000. Detects a band of approximately 45 kDa (predicted molecular weight: 48 kDa). |
IHC-P
1/2500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. See IHC antigen retrieval protocols. For unpurified use at 1/50 - 1/100. |
Target
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Function
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. -
Tissue specificity
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. -
Pathway
Protein modification; protein ubiquitination. -
Involvement in disease
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. -
Sequence similarities
Belongs to the WD repeat DDB2/WDR76 family.
Contains 5 WD repeats. -
Domain
The DWD box is required for interaction with DDB1. -
Post-translational
modificationsPhosphorylation 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. -
Cellular localization
Nucleus. Accumulates at sites of DNA damage following UV irradiation. - Information by UniProt
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Database links
- Entrez Gene: 1643 Human
- Omim: 600811 Human
- SwissProt: Q92466 Human
- Unigene: 700338 Human
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Alternative names
- damage-specific DNA binding protein 2 antibody
- Damage-specific DNA-binding protein 2 antibody
- DDB p48 subunit antibody
see all
Images
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Lane 1 : Anti-DDB2 antibody [EPR9811] (ab181136) at 1/2000 dilution
Lane 2 : Anti-DDB2 antibody [EPR9811] (ab181136) at 1/2000 dilution (Purified)
Lane 1 : SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysate
Lane 2 : Raji (Human Burkitt's lymphoma B lymphocyte) whole cell lysate
Lysates/proteins at 15 µg per lane.
Secondary
Lane 1 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Lane 2 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 48 kDa40kDa band might be an isomer of DNA damage-binding protein 2 (Isomer D3).
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Anti-DDB2 antibody [EPR9811] (ab181136) at 1/1000 dilution + HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 15 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Predicted band size: 48 kDa40kDa band might be an isomer of DNA damage-binding protein 2 (Isomer D3).
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All lanes : Anti-DDB2 antibody [EPR9811] (ab181136) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : DDB2 knockout HeLa cell lysate
Lane 3 : Raji cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 48 kDa
Observed band size: 48 kDaLanes 1-3: Merged signal (red and green). Green - ab181136 observed at 48 kDa. Red - loading control, ab8245 observed at 37 kDa.
ab181136 Anti-DDB2 antibody [EPR9811] was shown to specifically react with DDB2 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265115 (knockout cell lysate ab257177) was used. Wild-type and DDB2 knockout samples were subjected to SDS-PAGE. ab181136 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human stomach carcinoma tissue sections labeling DDB2 with Purified ab181136 at 1/2500 dilution (0.25 µg/mL). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (9)
ab181136 has been referenced in 9 publications.
- Kumar N et al. Global and transcription-coupled repair of 8-oxoG is initiated by nucleotide excision repair proteins. Nat Commun 13:974 (2022). PubMed: 35190564
- Jones TM et al. Targeted CUL4A inhibition synergizes with cisplatin to yield long-term survival in models of head and neck squamous cell carcinoma through a DDB2-mediated mechanism. Cell Death Dis 13:350 (2022). PubMed: 35428778
- Dardare J et al. DDB2 represses epithelial-to-mesenchymal transition and sensitizes pancreatic ductal adenocarcinoma cells to chemotherapy. Front Oncol 12:1052163 (2022). PubMed: 36568213
- Reichermeier KM et al. PIKES Analysis Reveals Response to Degraders and Key Regulatory Mechanisms of the CRL4 Network. Mol Cell 77:1092-1106.e9 (2020). PubMed: 31973889
- Steurer B et al. Fluorescently-labelled CPD and 6-4PP photolyases: new tools for live-cell DNA damage quantification and laser-assisted repair. Nucleic Acids Res 47:3536-3549 (2019). PubMed: 30698791
- Füzesi-Levi MG et al. CSNAP, the smallest CSN subunit, modulates proteostasis through cullin-RING ubiquitin ligases. Cell Death Differ N/A:N/A (2019). PubMed: 31367012
- Jang S et al. Damage sensor role of UV-DDB during base excision repair. Nat Struct Mol Biol 26:695-703 (2019). PubMed: 31332353
- Sun X et al. Senescence-associated secretory factors induced by cisplatin in melanoma cells promote non-senescent melanoma cell growth through activation of the ERK1/2-RSK1 pathway. Cell Death Dis 9:260 (2018). PubMed: 29449532
- Huang S et al. DDB2 Is a Novel Regulator of Wnt Signaling in Colon Cancer. Cancer Res 77:6562-6575 (2017). PubMed: 29021137