Anti-DDB2 antibody [2246C4a] (ab51017)
Key features and details
- Mouse monoclonal [2246C4a] to DDB2
- Suitable for: IHC-P, ICC/IF, WB, Flow Cyt
- Reacts with: Human
- Isotype: IgG1
Overview
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Product name
Anti-DDB2 antibody [2246C4a]
See all DDB2 primary antibodies -
Description
Mouse monoclonal [2246C4a] to DDB2 -
Host species
Mouse -
Tested applications
Suitable for: IHC-P, ICC/IF, WB, Flow Cytmore details -
Species reactivity
Reacts with: Human -
Immunogen
Recombinant fragment (N-terminal) Human.
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Positive control
- HeLa whole cell lysate
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General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at 4°C (stable for up to 12 months). Store at -20°C or -80°C. -
Storage buffer
pH: 7.40
Preservative: 0.05% Sodium azide
Constituents: 1% BSA, PBS -
Concentration information loading...
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Purity
Protein G purified -
Purification notes
Purified using protein G column chromatography from culture supernatant of hybridoma cultured in a medium containing bovine IgG depleted (approximately 95%) fetal bovine serum. Filtered through a 0.22 micrometer membrane. -
Clonality
Monoclonal -
Clone number
2246C4a -
Isotype
IgG1 -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
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Positive Controls
Applications
The Abpromise guarantee
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.
Application | Abreviews | Notes |
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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.
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ICC/IF | (1) |
Use a concentration of 10 µg/ml.
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WB | (1) |
1/50. Detects a band of approximately 45 kDa (predicted molecular weight: 48 kDa).
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Flow Cyt |
Use 1µg for 106 cells.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
Notes |
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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). |
Flow Cyt
Use 1µg for 106 cells. ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
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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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.
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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.
Protocols
Datasheets and documents
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Datasheet download
References (18)
ab51017 has been referenced in 18 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
- Shearer RF et al. K27-linked ubiquitylation promotes p97 substrate processing and is essential for cell proliferation. EMBO J 41:e110145 (2022). PubMed: 35349166
- Dai W et al. XPA Enhances Temozolomide Resistance of Glioblastoma Cells by Promoting Nucleotide Excision Repair. Cell Transplant 31:9636897221092778 (2022). PubMed: 35536165
- Zhu F et al. The miRNA125a-5p and miRNA125b-1-5p cluster induces cell invasion by down-regulating DDB2-reduced epithelial-to-mesenchymal transition (EMT) in colorectal cancer. J Gastrointest Oncol 13:3112-3122 (2022). PubMed: 36636074
- Zhang X et al. The CtBP1-HDAC1/2-IRF1 transcriptional complex represses the expression of the long noncoding RNA GAS5 in human osteosarcoma cells. Int J Biol Sci 15:1460-1471 (2019). PubMed: 31337976