Invitrogen

XPF Monoclonal Antibody (51)

Advanced Verification

This Antibody was verified by Relative expression to ensure that the antibody binds to the antigen stated. View Details

1 Published Figure

11 References

View all (33) XPF antibodies

Datasheet

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Datasheet

Protocols

Questions & Answers

Tech Support

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XPF Antibody in Western Blot (WB) — Western blot analysis was performed on nuclear extracts (30 µg lysate) of A-375 (Lane 1), A-431 (Lane 2), SK-OV-3 (Lane 3), Sf9 (Lane 4), Hep G2 (Lane 5), HeLa (Lane 6), A549 (Lane 7) and Jurkat (Lane 8). The blot was probed with Anti-XPF Mouse Monoclonal Antibody (Product # MA5-12060, 2 µg/mL) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal Secondary Antibody, HRP conjugate (Product # A28177, 0.4 µg/mL, 1:2500 dilution). A 104 kDa band corresponding to XPF was observed across the cell lines tested. Known quantity of protein samples were electrophoresed using Novex®NuPAGE®4-12 % Bis-Tris gel (Product # NP0321BOX), XCell SureLock Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

XPF Antibody in ChIP assay (ChIP) — Enrichment of endogenous XPF protein at specific gene loci using Anti-XPF Antibody: Chromatin Immunoprecipitation (ChIP) was performed using Anti-XPF Mouse Monoclonal Antibody (Product # MA5-12060, 3 µg) on sheared chromatin from 2 million HeLa cells treated with UV (2 hours) using the MAGnify ChIP system kit (Product # 49-2024). Normal Rabbit IgG was used as a negative IP control. The purified DNA was analyzed by qPCR with PCR primer pairs over EE1F1A, GAPDH promoter (active) and GAPDH gene, SAT alpha (inactive). Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method.

XPF Antibody — Figure 2 Detection and quantification of ERCC1/XPF foci numbers per nucleus (PLA) in A2780 ovarian cancer cells, untreated and after DDP. Nuclei of A2780 cells are shown in blue (DAPI) and ERCC1/XPF foci in the nuclei as green dots (FITC); magnification 40x, fluorescent microscopy. ( A ): ERCC1/XPF complexes expressed in untreated cells; ( B ): ERCC1/XPF foci in A2780 cells 24 h and 48 h after 10 muM (IC 50 ) and 20 uM DDP-treatment; ( C ): quantification of ERCC1/XPF foci number per nucleus in A2780 untreated and DDP-treated cells with 10 uM (IC 50 ) and 20 uM, fixed at two different time points. Each value is the mean +- standard deviation of three replicates and at least 50 nuclei were analyzed for each sample in different fields. Unpaired t -test was used to compare untreated vs. treated cells; * p <= 0.05.

Product Details

Applications

Tested Dilution

Publications

Western Blot (WB)

1-3 µg/mL

View 6 publications 6 publications

Immunocytochemistry (ICC/IF)

View 3 publications 3 publications

ChIP assay (ChIP)

1.5 µg/10^6 cells

Neutralization (Neu)

View 1 publication 1 publication

in situ PLA (PLA)

View 1 publication 1 publication

Product Specifications

Species Reactivity

Human

Published species

Human

Host/Isotype

Mouse / IgG2b, kappa

Class

Monoclonal

Type

Antibody

Clone

Immunogen

Recombinant human XPF protein

Conjugate

Unconjugated

Form

Liquid

Concentration

0.2 mg/mL

Purification

Protein A

Storage buffer

PBS, pH 7.4, with 0.2% BSA

Contains

0.09% sodium azide

Storage conditions

4° C

RRID

AB_10987557

Product Specific Information

MA5-12060 targets XPF in WB applications and shows reactivity with Human samples.

The MA5-12060 immunogen is recombinant human XPF protein.

Target Information

The structure specific ERCC 1 / XPF endonuclease complex is implicated in the repair of two distinct types of lesions in DNA: NER for UV induced lesions and bulky chemical adducts; and recombination repair of the very genotoxic interstrand cross links. NER mechanism involves dual incisions on both sides of the damage catalyzed by two nucleases. In mammalian cells, XPG cleaves 3' of the DNA lesion while the ERCC1 / XPF complex makes the 5' incision. ERCC1 / XPF probably plays a role in processing partially processed recombination intermediates.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Bioinformatics

Protein Aliases: DNA excision repair protein ERCC-4; DNA repair endonuclease XPF; DNA repair protein complementing XP-F cells; excision repair cross-complementation group 4; excision repair cross-complementing rodent repair deficiency, complementation group 4; excision-repair, complementing defective, in Chinese hamster; Xeroderma pigmentosum group F-complementing protein; xeroderma pigmentosum, complementation group F

Gene Aliases: ERCC11; ERCC4; FANCQ; RAD1; XFEPS; XPF

UniProt ID: (Human) Q92889

Entrez Gene ID: (Human) 2072

Molecular Function: DNA metabolism protein endodeoxyribonuclease nucleic acid metabolism protein