Invitrogen

ERCC1 Polyclonal Antibody

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ERCC1 Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of ERCC1 was performed using 70% confluent log phase U-87 MG cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with ERCC1 Polyclonal Antibody (Product # PA5-77971) at 1:100 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then with Goat anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32731) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b: Blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nuclear localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

ERCC1 Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — ERCC1 Polyclonal Antibody detects ERCC1 protein at cytoplasm and nucleus on human lung carcinoma by immunohistochemical analysis. Sample: Paraffin-embedded human lung carcinoma. ERCC1 Polyclonal Antibody (Product # PA5-77971) diluted at 1:250. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.

ERCC1 Antibody in Western Blot (WB) — Western blot was performed using Anti-ERCC1 Polyclonal Antibody (Product # PA5-77971) and a 36 kDa band corresponding to ERCC1 was observed across the cell lines tested. An uncharacterized band of ~ 17 kDa was also observed in U-87 MG and A-431 cell lines. Modified whole cell extracts (1% SDS) (30 µg lysate) of U-87 MG (Lane1), Hep G2 (Lane 2), HeLa (Lane 3) and A-431 (Lane 4) were electrophoresed using Novex® NuPAGE™ 10% Bis-Tris Protein Gel (Product # NP0302BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Product Details

PA5-77971

Applications	Tested Dilution	Publications
Western Blot (WB)	1:500-1:3,000	-
Immunohistochemistry (Paraffin) (IHC (P))	1:100-1:1,000	-
Immunocytochemistry (ICC/IF)	1:100-1:1,000	-

Product Specifications
Species Reactivity	Human, Rat
Host/Isotype	Rabbit / IgG
Class	Polyclonal
Type	Antibody
Immunogen	Recombinant protein encompassing a sequence within the center region of human ERCC1. The exact sequence is proprietary.
Conjugate	Unconjugated
Form	Liquid
Concentration	1.0 mg/mL
Purification	Antigen affinity chromatography
Storage buffer	PBS, pH 7, with 20% glycerol
Contains	0.025% Proclin 300
Storage conditions	Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
RRID	AB_2735768

Product Specific Information

Positive Control: 293T, A431, HeLa, HepG2, PC-12, Rat2

Predicted Reactivity: Mouse (86%), Pig (87%), Chimpanzee (100%), Bovine (87%)

Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.

Target Information

ERCC1 is a non-catalytic component of a structure-specific DNA repair endonuclease responsible for the 5'-incision during DNA repair. It is responsible, in conjunction with SLX4, for the first step in the repair of interstrand cross-links (ICL). It participates in the processing of anaphase bridge-generating DNA structures, which consist in incompletely processed DNA lesions arising during S or G2 phase, and can result in cytokinesis failure. ERCC1 is also required for homology-directed repair (HDR) of DNA double-strand breaks, in conjunction with SLX4. (Uniprot)

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

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Bioinformatics

Protein Aliases: DNA excision repair protein ERCC-1; excision repair cross-complementation group 1; excision repair cross-complementing 1; excision repair cross-complementing rodent repair deficiency, complementation group 1; excision repair cross-complementing rodent repair deficiency, complementation group 1 (includes overlapping antisense sequence)

Gene Aliases: COFS4; ERCC1; RAD10; UV20

UniProt ID: (Human) P07992

Entrez Gene ID: (Human) 2067, (Rat) 292673

Molecular Function: endodeoxyribonuclease