Isocitrate dehydrogenase [NADP]; D2HGA2; ICD-M; IDH; IDH2; IDHM; IDHP_HUMAN; IDP; IDPM; Isocitrate dehydrogenase [NADP], mitochondrial; Isocitrate dehydrogenase 2 (NADP+), mitochondrial; mNADP-IDH; NADP(+)-specific ICDH; Oxalosuccinate decarboxylase; isocitrate dehydrogenase 2 (NADP+), mitochondrial

For Research Use Only. Not for use in diagnostic procedures.

Polyclonal

Immunogen Affinity Purified

Lyophilized. Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.

At -20 degree C for one year. After reconstitution, at 4 degree C for one month. It can also be aliquoted and stored frozen at -20 degree C for a longer time. Avoid repeated freezing and thawing.

Manufactured in an ISO 13485:2003 and EN ISO 13485:2012 Certified Laboratory.

Small volumes of anti-IDH2 antibody vial(s) may occasionally become entrapped in the seal of the product vial during shipment and storage. If necessary, briefly centrifuge the vial on a tabletop centrifuge to dislodge any liquid in the container`s cap. Certain products may require to ship with dry ice and additional dry ice fee may apply.

Description: Rabbit IgG polyclonal antibody for Isocitrate dehydrogenase [NADP], mitochondrial(IDH2) detection. Tested with WB, IHC-P in Human; Mouse;Rat.

Background: Isocitrate dehydrogenase [NADP], mitochondrial is an enzyme that in humans is encoded by the IDH2 gene. Isocitrate dehydrogenases catalyze the oxidative decarboxylation of isocitrate to 2-oxoglutarate. These enzymes belong to two distinct subclasses, one of which utilizes NAD(+) as the electron acceptor and the other NADP(+). Five isocitrate dehydrogenases have been reported: three NAD(+)-dependent isocitrate dehydrogenases, which localize to the mitochondrial matrix, and two NADP(+)-dependent isocitrate dehydrogenases, one of which is mitochondrial and the other predominantly cytosolic. Each NADP(+)-dependent isozyme is a homodimer. The protein encoded by this gene is the NADP(+)-dependent isocitrate dehydrogenase found in the mitochondria. It plays a role in intermediary metabolism and energy production. This protein may tightly associate or interact with the pyruvate dehydrogenase complex. Alternative splicing results in multiple transcript variants.

Western Blot (WB), Immunohistochemistry (IHC) Paraffin

Western Blot Concentration: 0.1-0.5ug/ml
Immunohistochemistry (IHC) Paraffin Concentration: 0.5-1ug/ml

Figure 1. Western blot analysis of IDH2 using anti-IDH2 antibody (MBS177657).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: Rat Cardiac Muscle Tissue Lysate,
Lane 2: Rat Liver Tissue Lysate,
Lane 3: NIH3T3 Whole Cell Lysate,
Lane 4: SW620 Whole Cell Lysate,
Lane 5: HELA Whole Cell Lysate,
Lane 6: MCF-7 Whole Cell Lysate,
Lane 7: 22RV1 Whole Cell Lysate.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-IDH2 antigen affinity purified polyclonal antibody at 0.5ug/mL overnight at 4 degree C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for IDH2 at approximately 51KD. The expected band size for IDH2 is at 51KD.

Figure 2. IHC analysis of IDH2 using anti-IDH2 antibody (MBS177657).
IDH2 was detected in paraffin-embedded section of Mouse Intestine Tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml rabbit anti-IDH2 Antibody (MBS177657) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

Figure 3. IHC analysis of IDH2 using anti-IDH2 antibody (MBS177657).
IDH2 was detected in paraffin-embedded section of Rat Intestine Tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml rabbit anti-IDH2 Antibody (MBS177657) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

Figure 4. IHC analysis of IDH2 using anti-IDH2 antibody (MBS177657).
IDH2 was detected in paraffin-embedded section of Human Intestinal Cancer Tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml rabbit anti-IDH2 Antibody (MBS177657) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

Figure 5. IHC analysis of IDH2 using anti-IDH2 antibody (MBS177657).
IDH2 was detected in frozen section of human placenta tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml rabbit anti-IDH2 Antibody (MBS177657) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

Figure 6. IHC analysis of IDH2 using anti-IDH2 antibody (MBS177657).
IDH2 was detected in frozen section of mouse small intestine tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml rabbit anti-IDH2 Antibody (MBS177657) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

Figure 7. IHC analysis of IDH2 using anti-IDH2 antibody (MBS177657).
IDH2 was detected in frozen section of rat small intestine tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml rabbit anti-IDH2 Antibody (MBS177657) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

45,180 Da

isocitrate dehydrogenase (NADP(+)) 2, mitochondrial

isocitrate dehydrogenase [NADP], mitochondrial

Isocitrate dehydrogenase [NADP], mitochondrial

IDH??[Similar Products]

IDHP_HUMAN

Isocitrate dehydrogenases catalyze the oxidative decarboxylation of isocitrate to 2-oxoglutarate. These enzymes belong to two distinct subclasses, one of which utilizes NAD(+) as the electron acceptor and the other NADP(+). Five isocitrate dehydrogenases have been reported: three NAD(+)-dependent isocitrate dehydrogenases, which localize to the mitochondrial matrix, and two NADP(+)-dependent isocitrate dehydrogenases, one of which is mitochondrial and the other predominantly cytosolic. Each NADP(+)-dependent isozyme is a homodimer. The protein encoded by this gene is the NADP(+)-dependent isocitrate dehydrogenase found in the mitochondria. It plays a role in intermediary metabolism and energy production. This protein may tightly associate or interact with the pyruvate dehydrogenase complex. Alternative splicing results in multiple transcript variants. [provided by RefSeq, Feb 2014]

IDH2: a mitochondrial oxidoreductase that catalyzes the third step of the citric acid cycle: the oxidative decarboxylation of isocitrate, consuming NADP(+), and producing alpha-ketoglutarate (alpha-KG) and CO2. Can also use oxalosuccinate as a substrate. Alpha-KG is an activator the dioxygenases that hydroxylate the transcription factor HIF and lead to its degradation by VHL. Since HIF turns on oncogenic pathways, IDH2 has apparent tumor suppressor activity. Homo-dimerization is required for activity. Each subunit binds 1 magnesium or manganese ion. May tightly associate or interact with the pyruvate dehydrogenase complex. A somatic mutation resulting in the R172K conversion results in a neomorphic enzyme activity that converts isocitrate to R(-)-2-hydroxyglutarate (2HG) occurs in a significant percentage of patients with cytogenetically normal acute myeloid leukemia (AML). Elevated levels of 2HG are also correlated with an elevated risk of malignant brain tumors.

Protein type: EC 1.1.1.42; Other Amino Acids Metabolism - glutathione; Mitochondrial; Oxidoreductase; Carbohydrate Metabolism - citrate (TCA) cycle

Chromosomal Location of Human Ortholog: 15q26.1

Cellular Component: cytosol; mitochondrial inner membrane; mitochondrial matrix; mitochondrion; peroxisome

Molecular Function: isocitrate dehydrogenase (NADP+) activity; magnesium ion binding; NAD binding

Biological Process: 2-oxoglutarate metabolic process; carbohydrate metabolic process; glyoxylate cycle; isocitrate metabolic process; tricarboxylic acid cycle

Disease: D-2-hydroxyglutaric Aciduria 2

1. "Entrez Gene: IDH2 isocitrate dehydrogenase 2 (NADP+), mitochondrial". 2. Xu X, Zhao J, Xu Z, Peng B, Huang Q, Arnold E, Ding J (Aug 2004). "Structures of human cytosolic NADP-dependent isocitrate dehydrogenase reveal a novel self-regulatory mechanism of activity". The Journal of Biological Chemistry 279 (32): 33946-57.

All of MyBioSource's Products are for scientific laboratory research purposes and are not for diagnostic, therapeutics, prophylactic or in vivo use. Through your purchase, you expressly represent and warrant to MyBioSource that you will properly test and use any Products purchased from MyBioSource in accordance with industry standards. MyBioSource and its authorized distributors reserve the right to refuse to process any order where we reasonably believe that the intended use will fall outside of our acceptable guidelines.

While every efforts were made to ensure the accuracy of the information provided in this datasheet, MyBioSource will not be liable for any omissions or errors contained herein. MyBioSource reserves the right to make changes to this datasheet at any time without prior notice.

It is the responsibility of the customer to report product performance issues to MyBioSource within 30 days of receipt of the product. Please visit our Terms & Conditions page for more information.