Invitrogen

FEN1 Polyclonal Antibody

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FEN1 Antibody in Immunofluorescence (IF) — Immunofluorescence analysis of FEN1 was performed using 70% confluent log phase HeLa 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 FEN1 Polyclonal Antibody (Product # PA5-27518) at 1:200 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then with Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant Secondary Antibody, Alexa Fluor 488 (Product # A27034) 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 staining in nucleus. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

FEN1 Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemical analysis of FEN1 in the nucleus of paraffin-embedded human breast carcinoma using a FEN1 polyclonal antibody (Product # PA5-27518).

FEN1 Antibody in Western Blot (WB) — Western blot was performed using Anti-FEN1 Polyclonal Antibody, (Product # PA5-27518) and a 45 kDa band corresponding to FEN1 was observed in the cell lines and tissue tested. Modified whole cell extracts (1%SDS) (30 µg lysate) of A-431 (Lane 1), HCT 116 (Lane 2), HeLa (Lane 3), Hep G2 (Lane 4) and Mouse Spleen (Lane 5) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX). 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 conjugate (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)..

FEN1 Antibody in Immunoprecipitation (IP) — PA5-27518 immunoprecipitates FEN1 protein in IP experiments. IP samples: Jurkat whole cell extract A. 30 µg Jurkat whole cell extract B. Control with 4 µg of preimmune Rabbit IgG C. Immunoprecipitation of FEN1 protein by 4 µg FEN1 antibody (Product # PA5-27518) on 10% SDS-PAGE.

Product Details

Applications

Tested Dilution

Publications

Immunocytochemistry (ICC)

1:100-1:1000

Immunofluorescence (IF)

1:100-1:1000

Immunohistochemistry (IHC)

Assay Dependent

Immunohistochemistry (Paraffin) (IHC (P))

1:100-1:1000

Immunoprecipitation (IP)

1:100-1:500

Western Blot (WB)

1:500-1:3000

in situ PLA (PLA)

Assay Dependent

Product Specifications

Species

Human, Mouse, Rat

Expression System

Rabbit / IgG

Class

Polyclonal

Type

Antibody

Immunogen

Recombinant fragment corresponding to a region within amino acids 169 and 380 of Human FEN1

Conjugate

Unconjugated

Form

Liquid

Concentration

0.72 mg/mL

Purification

Antigen affinity chromatography

Storage buffer

0.1M tris glycine, pH 7, with 10% glycerol

Contains

0.01% thimerosal

Storage conditions

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

RRID

AB_2544994

Product Specific Information

Recommended positive controls: HepG2, NIH-3T3, JC, BCL-1, Raw264.7, PC-12.

Predicted reactivity: Mouse (96%), Rat (95%), Chicken (81%), Sheep (94%), Rhesus Monkey (98%), Bovine (96%).

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

Target Information

FEN-1 (Flap Endonuclease and Five prime Exonuclease) is a 42 a protein that functions as an endonuclease that cleaves branched DNA structures containing single-stranded 5' flaps or as an exonuclease on DNA nicks and gaps. FEN-1 endonuclease and exonuclease activities are stimulated by an association with proliferating cell nuclear antigen (PCNA) in the nucleus. FEN-1 is believed to have important roles in DNA replication and repair and is required in cell-free DNA replication systems.

⚠WARNING: This product can expose you to chemicals including mercury, which is known to the State of California to cause birth defects or other reproductive harm. For more information go to www.P65Warnings.ca.gov.

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

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Bioinformatics

Protein Aliases: DNase IV; FEN-1; FEN-1 {ECO:0000255|HAMAP-Rule:MF_03140}; FEN1; fen1 {ECO:0000255|HAMAP-Rule:MF_03140}; Flap endonuclease 1; flap endonuclease 1 {ECO:0000255|HAMAP-Rule:MF_03140}; Flap structure-specific endonuclease 1; flap structure-specific endonuclease 1 {ECO:0000255|HAMAP-Rule:MF_03140}; hFEN-1; Maturation factor 1; maturation factor-1; MF1

Gene Aliases: AW538437; FEN-1; FEN1; MF1; RAD2

UniProt ID: (Human) P39748, (Rat) Q5XIP6

Entrez Gene ID: (Human) 2237, (Mouse) 14156, (Rat) 84490

Function(s)