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Purified anti-IDH2 Antibody

Pricing & Availability
Clone
KrMab-3 (See other available formats)
Regulatory Status
RUO
Other Names
Isocitrate Dehydrogenase 2, Oxalosuccinate Decarboxylase, ICD-M, IDP, MNADP-IDH
Isotype
Mouse IgG2b, κ
Ave. Rating
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Product Citations
publications
KrMab-3_PURE_IDH2_Antibody_1_080321.png
Whole cell extracts (15 µg protein) from the indicated cell lines were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with 0.25 µg/mL (1:2000 dilution) of purified anti-IDH2 antibody (clone KrMab-3) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP goat anti-mouse IgG antibody (Cat. No. 405306) at a 2.0 µg/mL dilution. Direct-Blot™ HRP anti-GAPDH antibody (Cat. No. 607904) was used as a loading control at a 1:50000 dilution (lower). Lane M: Molecular weight marker.
  • KrMab-3_PURE_IDH2_Antibody_1_080321.png
    Whole cell extracts (15 µg protein) from the indicated cell lines were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with 0.25 µg/mL (1:2000 dilution) of purified anti-IDH2 antibody (clone KrMab-3) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP goat anti-mouse IgG antibody (Cat. No. 405306) at a 2.0 µg/mL dilution. Direct-Blot™ HRP anti-GAPDH antibody (Cat. No. 607904) was used as a loading control at a 1:50000 dilution (lower). Lane M: Molecular weight marker.
  • KrMab-3_PURE_IDH2_Antibody_2_080321.png
    HepG2 cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.5% Triton X-100 for 10 minutes, and blocked with 5% FBS for 60 minutes. Cells were then intracellularly stained with 5.0 µg/mL of purified anti-IDH2 antibody (panel A) followed by incubation with Alexa Fluor® 594 goat anti-mouse IgG antibody (Cat. No. 405326) 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). The merged image is shown in panel C. Nuclei were counterstained with DAPI, and the image was captured with a 60X objective.
  • KrMab-3_PURE_IDH2_Antibody_3_080321.png
    Whole cell extracts (250 μg total protein) prepared from Jurkat cells were immunoprecipitated overnight with 2.5 μg of purified mouse IgG2b, k isotype ctrl antibody (Cat. No. 400302) or purified anti-IDH2 antibody, (clone KrMab-3). The resulting IP fractions and whole cell extract input (6%) were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with KrMab-3 and a light chain specific mouse secondary antibody. Lane M: Molecular weight marker.
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946001 25 µg $118
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946002 100 µg $293
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Description

Isocitrate dehydrogenase 2 (IDH2) is a mitochondrial enzyme that is directly involved in the TCA cycle. The primary role of IDH2 is to convert isocitrate to 2-ketoglutarate (α-KG). The mutated isoform of IDH2 has been reported in glioblastomas, AML, and other cancers. The gain of function mutation in IDH2 leads to the accumulation of oncometabolite 2-hydroxyglutarate (2HG), which competitively inhibits HIF Prolyl Hydroxylase 2 (PHD2). This results in the stabilization and accumulation of hypoxia-inducible factors (HIF). The downstream effects of this mutation promote epigenetic changes and impaired cellular differentiation.

Product Details
Technical data sheet

Product Details

Verified Reactivity
Human, Mouse
Antibody Type
Monoclonal
Host Species
Mouse
Immunogen
Synthetic peptide corresponding to residues surrounding Ala174 of human IDH2 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, IP - 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 - 0.5 µg/mL. For immunocytochemistry, a concentration range of 2.0 - 5.0 μg/mL is recommended. For immunoprecipitation, the suggested use of this reagent is 2.5 µg/test. It is recommended that the reagent be titrated for optimal performance for each application.

Application Notes

This clone was tested for ICC using 4% PFA-fixed HepG2 cells permeabilized with either methanol or Triton X-100. While staining was observed with both methods, Triton X-100 permeabilization resulted in stronger staining.

RRID
AB_2894530 (BioLegend Cat. No. 946001)
AB_2894530 (BioLegend Cat. No. 946002)

Antigen Details

Biology Area
Cell Biology
Molecular Family
Enzymes and Regulators, Mitochondrial Markers
Antigen References
  1. Yan H. 2009. N Engl J Med. 360(8):765-73.
  2. Figueroa M, et al. 2010. Cancer Cell. 18:553-67.
  3. Wise D, et al. 2011. Proc Natl Acad Sci U S A. 108:19611-6.
Gene ID
3418 View all products for this Gene ID
UniProt
View information about IDH2 on UniProt.org
Go To Top Version: 1    Revision Date: 08/03/2021

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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