FEN1 Mouse Monoclonal Antibody [A1B12]
Catalog# HA600025
FEN1 Mouse Monoclonal Antibody [A1B12]
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WB
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IHC-P
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FC
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Human
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Mouse
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Rat
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unconjugated
Overview
Product Name
FEN1 Mouse Monoclonal Antibody [A1B12]
Antibody Type
Mouse Monoclonal Antibody
Immunogen
Recombinant protein within human FEN1 aa 150-380.
Species Reactivity
Human, Mouse, Rat
Validated Applications
WB, IHC-P, FC
Molecular Weight
Predicted band size: 43 kDa
Positive Control
K-562 cell lysate, SH-SY5Y cell lysate, PC-12 cell lysate, NIH/3T3 cell lysate, human tonsil tissue, human liver carcinoma tissue, human breast carcinoma tissue, K562.
Conjugation
unconjugated
Clone Number
A1B12
RRID
Product Features
Form
Liquid
Concentration
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.
Storage Buffer
1*TBS (pH7.4), 1% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG1
Purification Method
Protein G affinity purified.
Application Dilution
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WB
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1:500-1:2,000
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IHC-P
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1:100-1:500
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FC
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1:50-1:100
Target
Function
Flap endonuclease 1 is an enzyme that in humans is encoded by the FEN1 gene. The protein encoded by this gene removes 5' overhanging "flaps" (or short sections of single stranded DNA that "hang off" because their nucleotide bases are prevented from binding to their complementary base pair—despite any base pairing downstream) in DNA repair and processes the 5' ends of Okazaki fragments in lagging strand DNA synthesis. Direct physical interaction between this protein and AP endonuclease 1 during long-patch base excision repair provides coordinated loading of the proteins onto the substrate, thus passing the substrate from one enzyme to another. The protein is a member of the XPG/RAD2 endonuclease family and is one of ten proteins essential for cell-free DNA replication. DNA secondary structure can inhibit flap processing at certain trinucleotide repeats in a length-dependent manner by concealing the 5' end of the flap that is necessary for both binding and cleavage by the protein encoded by this gene. Therefore, secondary structure can deter the protective function of this protein, leading to site-specific trinucleotide expansions.
Background References
1. Zeng X. et. al. FEN1 mediates miR-200a methylation and promotes breast cancer cell growth via MET and EGFR signaling. FASEB J. 2019 Oct;33(10):10717-10730.
2. Li JL. et. al. FEN1 inhibitor increases sensitivity of radiotherapy in cervical cancer cells. Cancer Med. 2019 Dec;8(18):7774-7780.
Subcellular Location
Nucleus, nucleolus, nucleoplasm; Mitochondrion.
Synonyms
DNase IV antibody
FEN-1 antibody
FEN1 antibody
FEN1_HUMAN antibody
Flap endonuclease 1 antibody
Flap structure specific endonuclease 1 antibody
Flap structure-specific endonuclease 1 antibody
hFEN-1 antibody
hFEN1 antibody
Maturation factor 1 antibody
ExpandImages
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Western blot analysis of FEN1 on different lysates with Mouse anti-FEN1 antibody (HA600025) at 1/1,000 dilution.
Lane 1: K-562 cell lysate
Lane 2: SH-SY5Y cell lysate
Lysates/proteins at 10 µg/Lane.
Predicted band size: 43 kDa
Observed band size: 45 kDa
Exposure time: 10 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA600025) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature. -
Western blot analysis of FEN1 on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (HA600025, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:5,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: PC-12 cell lysate
Lane 2: NIH/3T3 cell lysate -
Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-FEN1 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA600025, 1/400) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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Immunohistochemical analysis of paraffin-embedded human liver carcinoma tissue using anti-FEN1 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA600025, 1/400) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using anti-FEN1 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA600025, 1/400) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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Flow cytometric analysis of FEN1 was done on K562 cells. The cells were fixed, permeabilized and stained with the primary antibody (HA600025, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Mouse IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"