Key Product Details

Species Reactivity

Validated:

Human, Mouse, Rat

Cited:

Human, Mouse

Predicted:

Guinea Pig (100%), Hamster (100%), Primate (100%), Rabbit (100%). Backed by our 100% Guarantee.

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Immunocytochemistry/ Immunofluorescence

Cited:

Western Blot, IF/IHC

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free
View all IDH2 Primary Antibodies »

Product Specifications

Immunogen

A synthetic peptide made to a C-terminal portion of the human IDH2 protein (between residues 375-452) [UniProt P48735].

Localization

Mitochondria

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for IDH2 Antibody - BSA Free

Western Blot: IDH2 AntibodyBSA Free [NBP2-22166]

Western Blot: IDH2 AntibodyBSA Free [NBP2-22166]

Western Blot: IDH2 Antibody [NBP2-22166] - 1) Non transfected 293T cells. 2) IDH2-flag (c-term) transfected 293T cells. WB image submitted by a verified customer review.
Immunocytochemistry/ Immunofluorescence: IDH2 Antibody - BSA Free [NBP2-22166]

Immunocytochemistry/ Immunofluorescence: IDH2 Antibody - BSA Free [NBP2-22166]

Immunocytochemistry/Immunofluorescence: IDH2 Antibody [NBP2-22166] - 293T cells transfected with IDH2-flag expression construct were fixed and stained with anti-IDH2 antibody followed by Alexa Fluor 488 anti-rabbit secondary antibody. DAPI staining shows nuclei. ICC/IF image submitted by a verified customer review.
Immunohistochemistry: IDH2 Antibody - BSA Free [NBP2-22166]

Immunohistochemistry: IDH2 Antibody - BSA Free [NBP2-22166]

Immunohistochemistry: IDH2 Antibody [NBP2-22166] - Staining of IDH2 in mouse intestine.
Western Blot: IDH2 AntibodyBSA Free [NBP2-22166]

Western Blot: IDH2 AntibodyBSA Free [NBP2-22166]

Western Blot: IDH2 Antibody [NBP2-22166] - Analysis of Caki-1 cell lysate (A), human skeletal muscle tissue (B), mouse skeletal muscle tissue (C), and rat skeletal muscle tissue (D) using IDH2 antibody at a concentration of 2 ug/mL.
Immunocytochemistry/ Immunofluorescence: IDH2 Antibody - BSA Free [NBP2-22166]

Immunocytochemistry/ Immunofluorescence: IDH2 Antibody - BSA Free [NBP2-22166]

Immunocytochemistry/Immunofluorescence: IDH2 Antibody [NBP2-22166] - IDH2 antibody was tested in U2OS cells with DyLight (green). Nuclei and alpha-tubulin were counterstained with DAPI (blue) and DyLight 550 (red).

Applications for IDH2 Antibody - BSA Free

Application
Recommended Usage

Immunocytochemistry/ Immunofluorescence

1:250 - 1:500

Immunohistochemistry

1:100 - 1:500

Immunohistochemistry-Paraffin

1:100 - 1:500

Western Blot

1 - 2 ug/mL
Application Notes
In Western blot a band is observed ~48 kDa. Prior to immunostaining paraffin tissues, antigen retrieval with sodium citrate buffer (pH 6.0) is recommended.

Reviewed Applications

Read 2 reviews rated 5 using NBP2-22166 in the following applications:

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS

Format

BSA Free

Preservative

0.02% Sodium Azide

Concentration

1.0 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: IDH2

Isocitrate dehydrogenase 2, also known as IDH, IDP, oxalosuccinate decarboxylase, IDPM, and mNADP-IDH, catalyzes the oxidative decarboxylation of isocitrate to 2-oxoglutarate. IDH2 is imperative for metabolic regulation and energy production, primarily via the citric acid cycle. In addition, IDH2 has been implicated in tight interactions with the pyruvate dehydrogenase complex. IDH2 is localized in the mitochondria and is found in stems, roots and leaves. Recent research has found that mutations in IDH2 can block histone demethylation and result in an inability for cells to differentiate in gliomas (PMID: 22343901). Further studies have also linked IDH2 to both gallbladder and bile duct cancer (PMID: 22180306).

Alternate Names

D2HGA2, EC 1.1.1.42, ICD-M, IDH, IDHM, IDP, IDPM, isocitrate dehydrogenase [NADP], mitochondrial, isocitrate dehydrogenase 2 (NADP+), mitochondrial, mNADP-IDH, NADP(+)-specific ICDH, Oxalosuccinate decarboxylase

Entrez Gene IDs

3418 (Human); 269951 (Mouse); 361596 (Rat)

Gene Symbol

IDH2

UniProt

P48735 (Human), P54071 (Mouse), P56574 (Rat)

Additional IDH2 Products

Product Documents for IDH2 Antibody - BSA Free

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

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Product Specific Notices for IDH2 Antibody - BSA Free

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Citations for IDH2 Antibody - BSA Free

Customer Reviews for IDH2 Antibody - BSA Free (2)

5 out of 5
2 Customer Ratings
5 Stars
100%
4 Stars
0%
3 Stars
0%
2 Stars
0%
1 Stars
0%

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Customer Images

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  • IDH2 Antibody
    Name: Anonymous
    Application: Immunocytochemistry
    Sample Tested: 293T
    Species: Human
    Verified Customer | Posted 01/02/2020
    293T cells transfected with IDH2-flag expression construct were fixed and stained with anti-IDH2 antibody followed by alexa488 anti-rabbit secondary antibody. DAPI staining shows nuclei.
    IDH2 Antibody - BSA Free NBP2-22166
  • IDH2 Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: 293T
    Species: Human
    Verified Customer | Posted 01/02/2020
    1. Non transfected 2. IDH2-flag (c-term) transfected
    IDH2 Antibody - BSA Free NBP2-22166

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Protocols

View specific protocols for IDH2 Antibody - BSA Free (NBP2-22166):

IDH2 Antibody:
Immunocytochemistry Protocol

Culture cells to appropriate density in 35 mm culture dishes or 6-well plates.

1. Remove culture medium and add 10% formalin to the dish. Fix at room temperature for 30 minutes.
2. Remove the formalin and add ice cold methanol. Incubate for 5-10 minutes.
3. Remove methanol and add washing solution (i.e. PBS). Be sure to not let the specimen dry out. Wash three times for 10 minutes.
4. To block nonspecific antibody binding incubate in 10% normal goat serum from 1 hour to overnight at room temperature.
5. Add primary antibody at appropriate dilution and incubate at room temperature from 2 hours to overnight at room temperature.
6. Remove primary antibody and replace with washing solution. Wash three times for 10 minutes.
7. Add secondary antibody at appropriate dilution. Incubate for 1 hour at room temperature.
8. Remove antibody and replace with wash solution, then wash for 10 minutes. Add Hoechst 33258 to wash solution at 1:25,0000 and incubate for 10 minutes. Wash a third time for 10 minutes.
9. Cells can be viewed directly after washing. The plates can also be stored in PBS containing Azide covered in Parafilm (TM). Cells can also be cover-slipped using Fluoromount, with appropriate sealing.

*The above information is only intended as a guide. The researcher should determine what protocol best meets their needs. Please follow safe laboratory procedures.

IDH2 Antibody:
Immunohistochemistry-Paraffin Embedded Sections

Antigen Unmasking:
Bring slides to a boil in 10 mM sodium citrate buffer (pH 6.0) then maintain at a sub-boiling temperature for 10 minutes. Cool slides on bench-top for 30 minutes.

Staining:
1. Wash sections in deionized water three times for 5 minutes each.
2. Wash sections in wash buffer for 5 minutes.
3. Block each section with 100-400 ul blocking solution for 1 hour at room temperature.
4. Remove blocking solution and add 100-400 ul diluted primary antibody. Incubate overnight at 4 C.
5. Remove antibody solution and wash sections in wash buffer three times for 5 minutes each.
6. Add 100-400 ul biotinylated diluted secondary antibody. Incubate 30 minutes at room temperature.
7. Remove secondary antibody solution and wash sections three times with wash buffer for 5 minutes each.
8. Add 100-400 ul Streptavidin-HRP reagent to each section and incubate for 30 minutes at room temperature.
9. Wash sections three times in wash buffer for 5 minutes each.
10. Add 100-400 ul DAB substrate to each section and monitor staining closely.
11. As soon as the sections develop, immerse slides in deionized water.
12. Counterstain sections in hematoxylin.
13. Wash sections in deionized water two times for 5 minutes each.
14. Dehydrate sections.
15. Mount coverslips.


IDH2 Antibody:
Western Blot Protocol

1. Perform SDS-PAGE on samples to be analyzed, loading 25 ug of total protein per lane.
2. Transfer proteins to membrane according to the instructions provided by the manufacturer of the membrane and transfer apparatus.
3. Stain according to standard Ponceau S procedure (or similar product) to assess transfer success, and mark molecular weight standards where appropriate.
4. Rinse the blot.
5. Block the membrane using standard blocking buffer for at least 1 hour.
6. Wash the membrane in wash buffer three times for 10 minutes each.
7. Dilute anti-IDH2 primary antibody in blocking buffer and incubate 1 hour at room temperature.
8. Wash the membrane in wash buffer three times for 10 minutes each.
9. Apply the diluted HRP conjugated secondary antibody in blocking buffer (as per manufacturers instructions) and incubate 1 hour at room temperature.
10. Wash the blot in wash buffer three times for 10 minutes each (this step can be repeated as required to reduce background).
11. Apply the detection reagent of choice in accordance with the manufacturers instructions.
Note: Tween-20 can be added to the blocking or antibody dilution buffer at a final concentration of 0.05-0.2%.

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