Invitrogen

XPF Monoclonal Antibody (51)

Advanced Verification

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

2 Published Figures

11 References

View all (33) XPF antibodies

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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 in Immunocytochemistry, Proximity Ligation Assay (PLA) (ICC/IF, PLA) — 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.

XPF Antibody — Antibody specificity was demonstrated by detection of enrichment of the target protein at specific gene loci. Chromatin Immunoprecipitation (ChIP) was performed using Anti-XPF Monoclonal Antibody (Product # MA5-12060) using PCR primer pairs over EE1F1A, GAPDH promoter (active) and GAPDH gene, SAT alpha (inactive). {RE}

Product Details

MA5-12060

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	51
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.

Product References

Impact of ERCC1, XPF and DNA Polymerase β Expression on Platinum Response in Patient-Derived Ovarian Cancer Xenografts.

Cancers

Guffanti F,Alvisi MF,Caiola E,Ricci F,De Maglie M,Soldati S,Ganzinelli M,Decio A,Giavazzi R,Rulli E,Damia G

MA5-12060 was used in Proximity Ligation Assay (PLA) to show experimental evidence that the ERCC1/XPF complex increases after DDP treatment and this possibly better associates with the cancer cells' abilities to activate the NER pathway to repair platinum-induced damage than its basal level.

Mon Aug 24 00:00:00 EDT 2020

Impact of ERCC1, XPF and DNA Polymerase β Expression on Platinum Response in Patient-Derived Ovarian Cancer Xenografts.

Cancers

Guffanti F,Alvisi MF,Caiola E,Ricci F,De Maglie M,Soldati S,Ganzinelli M,Decio A,Giavazzi R,Rulli E,Damia G

Mon Aug 24 00:00:00 EDT 2020

Impact of ERCC1, XPF and DNA Polymerase β Expression on Platinum Response in Patient-Derived Ovarian Cancer Xenografts.

Cancers

Guffanti F,Alvisi MF,Caiola E,Ricci F,De Maglie M,Soldati S,Ganzinelli M,Decio A,Giavazzi R,Rulli E,Damia G

Mon Aug 24 00:00:00 EDT 2020

Persistent 3'-phosphate termini and increased cytotoxicity of radiomimetic DNA double-strand breaks in cells lacking polynucleotide kinase/phosphatase despite presence of an alternative 3'-phosphatase.

DNA repair

Chalasani SL,Kawale AS,Akopiants K,Yu Y,Fanta M,Weinfeld M,Povirk LF

MA5-12060 was used in Western Blotting to suggest that, despite presence of an alternative 3'-phosphatase, loss of PNKP significantly sensitizes cells to 3'-phosphate-terminated DSBs, due to a 3'-dephosphorylation defect.

Wed Aug 01 00:00:00 EDT 2018

Mutational signatures reveal the role of RAD52 in p53-independent p21-driven genomic instability.

Genome biology

Galanos P,Pappas G,Polyzos A,Kotsinas A,Svolaki I,Giakoumakis NN,Glytsou C,Pateras IS,Swain U,Souliotis VL,Georgakilas AG,Geacintov N,Scorrano L,Lukas C,Lukas J,Livneh Z,Lygerou Z,Chowdhury D,Sørensen CS,Bartek J,Gorgoulis VG

Published figure using XPF monoclonal antibody (Product # MA5-12060) in Western Blot

Fri Mar 16 00:00:00 EDT 2018

A small molecule screen identifies an inhibitor of DNA repair inducing the degradation of TFIIH and the chemosensitization of tumor cells to platinum.

Chemistry & biology

Alekseev S,Ayadi M,Brino L,Egly JM,Larsen AK,Coin F

MA5-12060 was used in Western Blotting to find that spironolactone promotes a rapid and reversible degradation of XPB, a subunit of transcription/repair factor TFIIH.

Thu Mar 20 00:00:00 EDT 2014

Xeroderma pigmentosum group F protein binds to Eg5 and is required for proper mitosis: implications for XP-F and XFE.

Genes to cells : devoted to molecular & cellular mechanisms

Tan LJ,Saijo M,Kuraoka I,Narita T,Takahata C,Iwai S,Tanaka K

MA5-12060 was used in western blot to study the association between Eg5 and Xeroderma pigmentosum group F protein and the significance for mitosis and the pathogenesis of XP-F and XFE

Thu Mar 01 00:00:00 EST 2012

Delayed c-Fos activation in human cells triggers XPF induction and an adaptive response to UVC-induced DNA damage and cytotoxicity.

Cellular and molecular life sciences : CMLS

Tomicic MT,Reischmann P,Rasenberger B,Meise R,Kaina B,Christmann M

MA5-12060 was used in western blot to study the role of delayed c-Fos activation in the adaptive response to UVC-induced DNA damage and cytotoxicity

Sun May 01 00:00:00 EDT 2011

Repair of laser-localized DNA interstrand cross-links in G1 phase mammalian cells.

The Journal of biological chemistry

Muniandy PA,Thapa D,Thazhathveetil AK,Liu ST,Seidman MM

MA5-12060 was used in immunocytochemistry to study the repair of laser-localized DNA interstrand cross-links in G1 phase mammalian cells

Fri Oct 09 00:00:00 EDT 2009

Sequential recruitment of the repair factors during NER: the role of XPG in initiating the resynthesis step.

The EMBO journal

Mocquet V,Lainé JP,Riedl T,Yajin Z,Lee MY,Egly JM

MA5-12060 was used in blocking or activating experiment to study the recruitment of repair factors during nucleotide excision repair and the initiating role played by XPG

Wed Jan 09 00:00:00 EST 2008

gamma-H2AX formation in response to interstrand crosslinks requires XPF in human cells.

DNA repair

Mogi S,Oh DH

MA5-12060 was used in western blot to examine the role of XPF

Sat Jun 10 00:00:00 EDT 2006

Phosphorylation of XPB helicase regulates TFIIH nucleotide excision repair activity.

The EMBO journal

Coin F,Auriol J,Tapias A,Clivio P,Vermeulen W,Egly JM

MA5-12060 was used in western blot to study the role of XPB helicase phosphorylation in the regulation of the nucleotide excision repair activity of TFIIH

Wed Dec 08 00:00:00 EST 2004

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