Invitrogen

FXN Polyclonal Antibody

Advanced Verification

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

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Datasheet

Protocols

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Tech Support

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FXN Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of Frataxin, mitochondrial was performed using 70% confluent log phase PC-3 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 45 minutes at room temperature. The cells were labeled with FXN Polyclonal Antibody (Product # PA5-95829) at 1:100 in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (1:2000), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with Hoechst 33342 (Product # H1399). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic(mitochondrial like pattern) localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 40X magnification in CellInsight CX7 LZR High-Content Screening (HCS) Platform (Product # CX7C1115LZR).

FXN Antibody in Western Blot (WB) — Western Blot was performed using Anti-FXN Polyclonal Antibody (Product # PA5-95829) and a 14 kDa band corresponding to Frataxin, mitochondrial was observed across cell lines tested. Membrane enriched extracts (30 µg lysate) of U-87 MG (Lane 1), Jurkat (Lane 2), A-431 (Lane 3), HeLa (Lane 4), PC-12 (Lane 5) and tissue extracts (30 µg lysate) of Mouse Kidney (Lane 6), Mouse Heart (Lane 7) and Rat Kidney (Lane 8) were electrophoresed using Novex™ 16% Tricine Protein Gel (Product # EC6695BOX). 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 (1:2000 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 SuperSignal™ West Dura Extended Duration Substrate (Product # 34076).

FXN Antibody — Antibody specificity was demonstrated by siRNA mediated knockdown of target protein. HeLa cells were transfected with Frataxin, mitochondrial siRNA and decrease in signal intensity was observed in Western Blot application using Anti-FXN Polyclonal Antibody (Product # PA5-95829). {KD}

Product Details

PA5-95829

Applications	Tested Dilution	Publications
Western Blot (WB)	1:500-1:2,000	-
Immunocytochemistry (ICC/IF)	1:100	-

Product Specifications
Species Reactivity	Human, Mouse, Rat
Host/Isotype	Rabbit / IgG
Class	Polyclonal
Type	Antibody
Immunogen	Recombinant fusion protein containing a sequence corresponding to amino acids 42-210 of human FXN / Frataxin View antigen
Conjugate	Unconjugated
Form	Liquid
Concentration	4.7 mg/mL
Purification	Antigen affinity chromatography
Storage buffer	PBS, pH 7.3, with 50% glycerol
Contains	0.02% sodium azide
Storage conditions	-20° C, Avoid Freeze/Thaw Cycles
RRID	AB_2807631

Product Specific Information

Immunogen sequence: LRTDIDATCT PRRASSNQRG LNQIWNVKKQ SVYLMNLRKS GTLGHPGSLD ETTYERLAEE TLDSLAEFFE DLADKPYTFE DYDVSFGSGV LTVKLGGDLG TYVINKQTPN KQIWLSSPSS GPKRYDWTGK NWVYSHDGVS LHELLAAELT KALKTKLDLS SLAYSGKDA; Positive Samples: K-562, U-87MG, Jurkat, Raji, DU-145, Mouse kidney, Mouse liver, Mouse heart; Cellular Location: Cytoplasm, Mitochondrion

Target Information

FXN is a mitochondrial protein which belongs to the FRATAXIN family. The protein functions in regulating mitochondrial iron transport and respiration. The expansion of intronic trinucleotide repeat GAA results in Friedreich ataxia.

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

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Bioinformatics

Protein Aliases: d-FXN; Frataxin intermediate form; Frataxin mature form; Frataxin(56-210); Frataxin(78-210); Frataxin(81-210); Frataxin, mitochondrial; Friedreich ataxia; Friedreich ataxia protein; Fxn; i-FXN; m56-FXN; m78-FXN; m81-FXN

Gene Aliases: CyaY; FA; FARR; FRDA; FXN; RGD1565754; X25

UniProt ID: (Human) Q16595, (Mouse) O35943

Entrez Gene ID: (Human) 2395, (Rat) 499335, (Mouse) 14297

Molecular Function: transfer/carrier protein