Key features and details
- Mouse monoclonal [2246C4a] to DDB2
- Suitable for: IHC-P, ICC/IF, WB, Dot blot, Flow Cyt
- Reacts with: Human
- Isotype: IgG1
Product nameAnti-DDB2 antibody [2246C4a]
See all DDB2 primary antibodies
DescriptionMouse monoclonal [2246C4a] to DDB2
Tested applicationsSuitable for: IHC-P, ICC/IF, WB, Dot blot, Flow Cytmore details
Species reactivityReacts with: Human
Recombinant fragment (N-terminal) Human.
- HeLa whole cell lysate
Storage instructionsShipped at 4°C. Store at 4°C (stable for up to 12 months). Store at -20°C or -80°C.
Storage bufferpH: 7.40
Preservative: 0.05% Sodium azide
Constituents: 1% BSA, PBS
Concentration information loading...
PurityProtein G purified
Purification notesPurified 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.
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.
|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.
FunctionRequired 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 specificityUbiquitously 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.
PathwayProtein modification; protein ubiquitination.
Involvement in diseaseDefects 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 similaritiesBelongs to the WD repeat DDB2/WDR76 family.
Contains 5 WD repeats.
DomainThe DWD box is required for interaction with DDB1.
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 localizationNucleus. Accumulates at sites of DNA damage following UV irradiation.
- Information by UniProt
- damage-specific DNA binding protein 2 antibody
- Damage-specific DNA-binding protein 2 antibody
- DDB p48 subunit antibody
Anti-DDB2 antibody [2246C4a] (ab51017) at 1/50 dilution + HeLa whole cell lysate at 50 µg
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.
ab51017 has been referenced in 14 publications.
- 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
- Kottemann MC et al. Removal of RTF2 from Stalled Replisomes Promotes Maintenance of Genome Integrity. Mol Cell 69:24-35.e5 (2018). PubMed: 29290612
- Piquet S et al. The Histone Chaperone FACT Coordinates H2A.X-Dependent Signaling and Repair of DNA Damage. Mol Cell 72:888-901.e7 (2018). PubMed: 30344095
- Zhu B et al. The protective role of DOT1L in UV-induced melanomagenesis. Nat Commun 9:259 (2018). PubMed: 29343685
- Rüthemann P et al. Chromatin remodeler CHD1 promotes XPC-to-TFIIH handover of nucleosomal UV lesions in nucleotide excision repair. EMBO J 36:3372-3386 (2017). PubMed: 29018037