Western blot analysis was performed on nuclear extracts (30 ug 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 ug/ml) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.4 ug/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 Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
|Tested species reactivity||Human|
|Published species reactivity||Human|
|Host / Isotype||Mouse / IgG2b, kappa|
|Immunogen||Recombinant human XPF protein|
|Storage buffer||PBS, pH 7.4, with 0.2% BSA|
|Contains||0.09% sodium azide|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Western Blot (WB)||1-3 µg/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
MA5-12060 targets XPF in WB applications and shows reactivity with Human samples.
The MA5-12060 immunogen is recombinant human XPF protein.
The structure-specific ERCC1/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.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Xeroderma pigmentosum group F protein binds to Eg5 and is required for proper mitosis: implications for XP-F and XFE.
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
|Tan LJ,Saijo M,Kuraoka I,Narita T,Takahata C,Iwai S,Tanaka K||Genes to cells : devoted to molecular and cellular mechanisms (17:173)||2012|
Delayed c-Fos activation in human cells triggers XPF induction and an adaptive response to UVC-induced DNA damage and cytotoxicity.
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
|Tomicic MT,Reischmann P,Rasenberger B,Meise R,Kaina B,Christmann M||Cellular and molecular life sciences : CMLS (68:1785)||2011|
gamma-H2AX formation in response to interstrand crosslinks requires XPF in human cells.
MA5-12060 was used in western blot to examine the role of XPF
|Mogi S,Oh DH||DNA repair (5:731)||2006|
Phosphorylation of XPB helicase regulates TFIIH nucleotide excision repair activity.
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
|Coin F,Auriol J,Tapias A,Clivio P,Vermeulen W,Egly JM||The EMBO journal (23:4835)||2004|
Repair of laser-localized DNA interstrand cross-links in G1 phase mammalian cells.
MA5-12060 was used in immunocytochemistry to study the repair of laser-localized DNA interstrand cross-links in G1 phase mammalian cells
|Muniandy PA,Thapa D,Thazhathveetil AK,Liu ST,Seidman MM||The Journal of biological chemistry (284:27908)||2009|
Sequential recruitment of the repair factors during NER: the role of XPG in initiating the resynthesis step.
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
|Mocquet V,Lainé JP,Riedl T,Yajin Z,Lee MY,Egly JM||The EMBO journal (27:155)||2008|