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Purified anti-Fumarase Antibody

Pricing & Availability
Clone
W18319C (See other available formats)
Regulatory Status
RUO
Other Names
Fumarate Hydratase, FH
Isotype
Rat IgG2a, κ
Ave. Rating
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Product Citations
publications
W18319C_PURE_Fumerase_Antibody_1_030921
Whole cell extracts (15 µg protein) from the indicated cell lines were resolved on a 4-12% Bis-Tris gel, transferred to PVDF and probed with 1.0 µg/mL (1:500 dilution) of purified anti-fumarase antibody (clone W18319C) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP goat anti-rat IgG antibody (Cat. No. 405405) at a 1:3000 dilution. Direct-Blot™ HRP anti-GAPDH antibody (Cat. No. 607904) was used as a loading control at a 1:25000 dilution (lower). Lane M: Molecular weight marker.
  • W18319C_PURE_Fumerase_Antibody_1_030921
    Whole cell extracts (15 µg protein) from the indicated cell lines were resolved on a 4-12% Bis-Tris gel, transferred to PVDF and probed with 1.0 µg/mL (1:500 dilution) of purified anti-fumarase antibody (clone W18319C) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP goat anti-rat IgG antibody (Cat. No. 405405) at a 1:3000 dilution. Direct-Blot™ HRP anti-GAPDH antibody (Cat. No. 607904) was used as a loading control at a 1:25000 dilution (lower). Lane M: Molecular weight marker.
  • W18319C_PURE_Fumerase_Antibody_2_030921
    HeLa cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with methanol for 10 minutes, and blocked with 5% FBS for 60 minutes. Cells were then intracellularly stained with 5.0 µg/mL of purified anti-fumarase antibody (clone W18319C) (panel A) followed by incubation with Alexa Fluor® 594 goat anti-rat IgG antibody (Cat. No. 405422) at 2.0 µg/mL. Alexa Fluor® 647 anti-Cytochrome c antibody (Cat. No. 612310) was used at 5.0 µg/mL to co-stain the mitochondria and is presented in green pseudocolor (panel B).
  • W18319C_PURE_Fumerase_Antibody_3_040323
    IHC staining of purified anti-Fumarase (clone W18319C) on formalin-fixed paraffin-embedded human kidney tissue. Following antigen retrieval using Tris-EDTA (10 mM Tris, 1 mM EDTA, pH 9.0), the tissue was incubated with 1.0 µg/mL of anti-Fumarase (clone W18319C) followed by incubation with 2.5 µg/mL of Alexa Fluor® 647 goat anti-rat IgG (Cat. No. 405416) for 1 hour at room temperature. Nuclei were counterstained with DAPI (Cat. No. 422801). Images were captured with a 10X (panel A) and 40X (panel B) objective. Scale bar: 20 µm
Cat # Size Price Quantity Check Availability Save
940001 25 µg £81
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940002 100 µg £201
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Description

Fumarate hydratase, or fumarase, is a Krebs cycle enzyme that catalyzes the conversion of fumarate to L-malate. A cytosolic isoform of Fumarase has been described that is required for arginine metabolism and the urea cycle. Inactivating germline mutations of the FH gene are found in patients with hereditary leiomyomatosis and renal cell cancer syndrome, which predisposes patients to increased risk of developing uterine and cutaneous leipmyomas and renal cell carcinomas. Mechanistically, elevated fumarate functions as an oncometabolite by inhibiting the activity of α-ketoglutarate dependent enzymes, such as hypoxia-inducible factor prolyl hydroxylases (PHDs) and ten-eleven translocation (TET) dioxygenases.

Product Details
Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Human, Mouse
Antibody Type
Monoclonal
Host Species
Rat
Immunogen
Partial recombinant human fumarase protein
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide
Preparation
The antibody was purified by affinity chromatography.
Concentration
0.5 mg/mL
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C.
Application

WB - Quality tested
ICC, IHC-P - Verified
Recommended Usage

Each lot of this antibody is quality control tested by western blotting. For western blotting, the suggested use of this reagent is 0.1 - 1.0 µg/mL. For immunocytochemistry, a concentration range of 1.0 - 5.0 μg/mL is recommended. For immunohistochemistry on formalin-fixed paraffin-embedded tissue sections, a concentration range of 1.0 - 10.0 µg/mL is suggested. It is recommended that the reagent be titrated for optimal performance for each application.

Application Notes

This clone was tested for ICC on HeLa cells fixed with 4% PFA and permeabilized with either methanol or Triton X-100. Both methods were compatible fumarase staining.

This clone failed to immunoprecipitate fumarase from HeLa extracts. We therefore do not recommend this clone for the application.

RRID
AB_2890856 (BioLegend Cat. No. 940001)
AB_2890856 (BioLegend Cat. No. 940002)

Antigen Details

Structure
Fumarase is a 510 amino acid protein with a predicted molecular weight of 55 kD.
Distribution

Ubiquitously expressed/Mitochondrial, Cytosolic

Interaction
H2AZ1
Biology Area
Cell Biology, Mitochondrial Function
Molecular Family
Enzymes and Regulators
Antigen References
  1. Laukka T, et al. 2016. J Biol Chem. 291:4256-65.
  2. Sciavoceli M, et al. 2016. Nature. 537:544-547.
Gene ID
2271 View all products for this Gene ID
UniProt
View information about Fumarase on UniProt.org
Go To Top Version: 2    Revision Date: 04/03/2023

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

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