This is a knockout-validated antibody summary, based on the publication "Repair protein persistence at DNA lesions characterizes XPF defect with Cockayne syndrome features", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

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Figure 1. (A) Immunoblot showing XPF and ERCC1 expression in U2OS, XPF knockout (KO) and XPF-GFP (XPF-wt) expressing XPF KO cells. Ku70 staining is shown as loading control. (B) Immunofluorescence pictures showing localization of endogenous XPF in U2OS (top panel) and XPF-wt in XPF KO cells (bottom panel) after UVC irradiation. Scale bar: 5 um. Please see Figure XXXX in the article [1].

Mouse monoclonal IgG1 (kappa light chain)

Company: Santa Cruz

Antibody: XPF

Catalog number: sc-136153

Summary: Mouse monoclonal IgG1 (kappa light chain) against His-tagged recombinant protein corresponding to amino acids 629-905 of human XPF. Recommended for detection of XPF of mouse, rat and human origin by western blot, immunoprecipitation and ELISA.

Western blot

Parental, XPF-knockout and XPF-GFP reconstituted U2OS cell lines.

WB: 2% BSA.

IC: PBS containing 0.15% glycine and 0.5% BSA.

WB: for 2 h or overnight.

IC: for 2 h.

WB: Secondary antibodies were conjugated with CF IRDye 680 or 770 (Sigma).

IC: Alexa Fluor conjugated secondary antibodies (488, 555 and 633; Invitrogen) for 1 h.

WB: Odyssey CLx Infrared Imaging System (LI-COR Biosciences).

IC: LSM700 microscope equipped with a 40x Plan-apochromat 1.3 NA oil immersion lens (Carl Zeiss).

The same clone (3F2/3) is sold as Santa Cruz Biotechnology sc-136153; Abcam ab85140.