The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 µg/ml.
1/2000 - 1/10000. Detects a band of approximately 39 kDa (predicted molecular weight: 31 kDa).
Use at 2-5 µg/mg of lysate.
1/200 - 1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Involved in DNA excision repair. Initiates repair by binding to damaged sites with various affinities, depending on the photoproduct and the transcriptional state of the region. Required for UV-induced CHK1 phosphorylation and the recruitment of CEP164 to cyclobutane pyrimidine dimmers (CPD), sites of DNA damage after UV irradiation.
Expressed in various cell lines and in skin fibroblasts.
Involvement in disease
Defects in XPA are a cause of xeroderma pigmentosum complementation group A (XP-A) [MIM:278700]; also known as xeroderma pigmentosum type 1 (XP1). XP-A 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. Group A patients show the most severe skin symptoms and progressive neurological disorders.
Belongs to the XPA family.
Phosphorylated upon DNA damage, probably by ATM or ATR. Ubiquitinated by HERC2 leading to degradation by the proteasome.
ICC/IF image of ab85914 stained HepG2 cells. The cells were 100% methanol fixed (5 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 (ab85914, 1µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 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.
Western blot - Anti-XPA antibody (ab85914)
All lanes : Anti-XPA antibody (ab85914) at 0.04 µg/ml
Lane 1 : HeLa whole cell lysate at 50 µg Lane 2 : HeLa whole cell lysate at 15 µg Lane 3 : HeLa whole cell lysate at 5 µg
Exposure time : 30 secondsDetection: Chemiluminescence with exposure time of 30 seconds
Immunoprecipitation - Anti-XPA antibody (ab85914)
Detection of XPA by Western Blot of Immunprecipitate.
ab85914 at 0.1µg/ml staining XPA in HeLa whole cell lysate immunoprecipitated using ab85914 at 3µg/mg lysate (1 mg/IP; 20% of IP loaded/lane). Detection: Chemiluminescence with exposure time of 30 seconds.
Zhou X et al. Differential binding of monomethylarsonous acid compared to arsenite and arsenic trioxide with zinc finger peptides and proteins. Chem Res Toxicol27:690-8 (2014).
Read more (PubMed: 24611629) »
King BS et al. Poly(ADP-ribose) contributes to an association between poly(ADP-ribose) polymerase-1 and xeroderma pigmentosum complementation group A in nucleotide excision repair. J Biol Chem287:39824-33 (2012).
Read more (PubMed: 23038248) »