Anti-XPA antibody, mouse monoclonal (5F12)
Product code70-031Size

100 µgStorage-20℃

Concentration

1.0 mg/ml

Buffer

PBS- with 50% glycerol

PurityPurified IgG fraction with protein A from hybridoma cell culture medium.

Immunogen

Recombinant full-length human XPA protein

Isotype

Mouse IgG2b

Reactivity

Human (expected to react also with mouse XPA from the sequence homology)

Special notes

Epitope: Amino acids 30-47

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 TFIIHOther applications have not been tested.

Background

XP (Xeroderma pigmentosum) is an autosomal recessive human diseasecharacterized 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 inearly step of NER. The hybridoma 5F12 was constructed by Prof. K. Tanaka’s
group who first cloned the XPA gene (2, 3).
Data Link

UniProtKB/Swiss-Prot P23025 (XPA_HUMAN)

Please note: All products are FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC
PROCEDURES. NOT FOR MILITARY USE.
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Data Images: 70-031 Anti-XPA antibody, mouse monoclonal (5F12)

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 Western 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.

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No UVUV (20 J/m2)

Fig.2 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 withparaform 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 whichinterefere 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

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