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No More Sales-Anti-XPA antibody, monoclonal (5F12) (250 ug)

Product#: inactived-FNK-70-032

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Anti-XPA antibody, monoclonal (5F12)

inactived-FNK-70-032

General information

Cat. No.	： inactived-FNK-70-032
Size	：250 µg
Antigen Species	：Human
Host Species	：Mouse
Immunogen	：Recombinant full-length human XPA protein
Clone No.	：5F12
Cross Reactivity	：Human (expected to react also with mouse XPA from the sequence homology)
Epitope	：Amino acids 30-47
Subtype	：IgG2b
Form	：Purified IgG, 1 mg/ml in PBS pH 7.2, 50% glycerol, filter-sterilized
Application	：1. Western blotting (1/1,000~1/10,000) ：2. Immunofluorescence staining (1/100~1/1,000) ：3. ELISA ：4. Inhibition of in vitro excision repair reaction ：5. Inhibition of XPA interaction with ERCC1 and TFIIH ：Other applications have not been tested.
Shipping and Storage	：Ship at 4℃ or -20℃, and store at -20℃. Do not freeze.
Data Link	：UniProtKB/Swiss-Prot P23025 (XPA_HUMAN)

Cat. No.	Size	Link
FNK-70-031	100 µg
FNK-70-032	250 µg

Antigen Characterization

Type DNA repair protein
Application DNA damage and repair studies, UV sensitivity assays
Species Human
Recombinant Yes

XPA in the Cells

XPA plays crucial rokes in the NER pathway

DNA damage verification
Recruitment of other NER proteins
Acts as a molecular scaffold to assemble NER factors around damage sites
Stabilization of NER intermediates

It also interacts with multiple proteins both within and outside the NER pathway which include:

ERCC1 (part of the DNA-incising complex)
Replication protein A1 (RPA1)
XAB2 (XPA-binding protein 2)

XPA in the Human Body

XP (Xeroderma pigmentosum) is an autosomal recessive human disease characterized by hypersensitivity to sunlight and a high incidence of skin cancer on sun-exposed skin (1). Cells from XP patients are hypersensitive to killing by UV irradiation because of a defect in nucleotide excision repair (NER). XP is classified into seven complementation groups (A~G) and a variant form (1). XPA shows the most severe symptoms. Products encoded by the XP genes function in repairing UV-induced cyclobutane pyrimidine dimmer and (6-4) photoproducts as well as chemically induced variety of DNA lesions (1). XPA protein consists of 273 amino acids and forms a complex with many proteins such as RPA, ERCC1, TFIIH、XAB1, and XAB2, which plays a role in early step of NER. The hybridoma 5F12 was constructed by Prof. K. Tanaka’s group who first cloned the XPA gene (2, 3).


Fig. 1 Western blot of endogenous XpA protein. Hela cell whole extrac (20 μg)t. Antibody was used at 1/2,000 dilution. As secondary antibody, HRP conjugated goat anti-mouse IgG was used at 1/20,000 dilution.	Fig 2 Weastern blot of XpA: Validation. Lane 1. Extract of Hela cells (XpA wild type) Lane 2. Extract of XP12ROSV cells (XpA deficient ) The primary and the secondary antibodies were used at 1/2,000 and 1/20,000 dilutions.

	Fig.3 Immunofluorescence staining of human fibroblast cells (GM0637) using anti-XpA antibody (5F12) The cells were non-irradiated (left) or irradiated with UV at 20 J/m2 and fixed after 30 min with paraform aldehyde. The antibody was used at 1/100 dilution and as the second antibody, Alexa 488 conjugated goat anti-mouse IgG was used at 1/5,000 dilution.

References

This antibody is described and used in Ref. 2

1. Friedberg EC et al DNA Repair and Mutagenesis 2nd ed., ASM Press (2006)
2. Saijo M et al “Inhibition of nucleotide excision repair by anti-XPA monoclonal antibodies which interefere with binding to RPA, ERCC1, and TFIIH” Biochem Biophys Res Comm 321:815-822 (2004) PMID: 15358100
3. Tanaka K et al “Analysis of a human DNA excision repair gene involved in group A xeroderma pigmentosum and containing a zinc-finger domain” Nature 348:73 -76 (1990) PMID: 2234061