Recombinant Anti-IDH2 antibody [EPR7577] - BSA and Azide free (ab230796)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR7577] to IDH2 - BSA and Azide free
- Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
-
Product name
Anti-IDH2 antibody [EPR7577] - BSA and Azide free
See all IDH2 primary antibodies -
Description
Rabbit monoclonal [EPR7577] to IDH2 - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IFmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
-
Positive control
- MOLT4, K562, U87-M, and HepG2 cell lysates; Human liver tissue
-
General notes
ab230796 is the carrier-free version of ab131263.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
Properties
-
Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Dissociation constant (KD)
KD = 4.70 x 10 -11 M Learn more about KD -
Storage buffer
pH: 7.20
Constituent: PBS -
Carrier free
Yes -
Concentration information loading...
-
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR7577 -
Isotype
IgG -
Research areas
Associated products
-
Alternative Versions
-
Compatible Secondaries
-
Conjugation kits
-
Isotype control
-
Positive Controls
-
Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab230796 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
---|---|---|
Flow Cyt (Intra) |
Use at an assay dependent concentration.
ab199376 - Rabbit monoclonal IgG (Low endotoxin, Azide free), is suitable for use as an isotype control with this antibody. |
|
WB |
Use at an assay dependent concentration. Detects a band of approximately 45 kDa (predicted molecular weight: 50 kDa).
|
|
IHC-P | (1) |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
|
ICC/IF |
Use at an assay dependent concentration.
|
Notes |
---|
Flow Cyt (Intra)
Use at an assay dependent concentration. ab199376 - Rabbit monoclonal IgG (Low endotoxin, Azide free), is suitable for use as an isotype control with this antibody. |
WB
Use at an assay dependent concentration. Detects a band of approximately 45 kDa (predicted molecular weight: 50 kDa). |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
ICC/IF
Use at an assay dependent concentration. |
Target
-
Function
Plays a role in intermediary metabolism and energy production. It may tightly associate or interact with the pyruvate dehydrogenase complex. -
Involvement in disease
D-2-hydroxyglutaric aciduria 2
Glioma
enetic variations are associated with cartilaginous tumors such as enchondroma or chondrosarcoma. -
Sequence similarities
Belongs to the isocitrate and isopropylmalate dehydrogenases family. -
Post-translational
modificationsAcetylation at Lys-413 dramatically reduces catalytic activity. Deacetylated by SIRT3. -
Cellular localization
Mitochondrion. - Information by UniProt
-
Database links
- Entrez Gene: 3418 Human
- Entrez Gene: 269951 Mouse
- Entrez Gene: 361596 Rat
- Omim: 147650 Human
- SwissProt: P48735 Human
- SwissProt: P54071 Mouse
- SwissProt: P56574 Rat
- Unigene: 596461 Human
see all -
Alternative names
- D2HGA2 antibody
- ICD-M antibody
- IDH antibody
see all
Images
-
All lanes : Anti-IDH2 antibody [EPR7577] (ab131263) at 1/1000 dilution
Lane 1 : Wild-type Jurkat cell lysate
Lane 2 : IDH2 knockout Jurkat cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 50 kDaFalse colour image of Western blot: Anti-IDH2 antibody [EPR7577] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab131263 was shown to bind specifically to IDH2. A band was observed at 48 kDa in wild-type Jurkat cell lysates with no signal observed at this size in IDH2 knockout cell line ab282331 (knockout cell lysate ab283148). To generate this image, wild-type and IDH2 knockout Jurkat cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
-
Lanes 1-7 : Anti-IDH2 antibody [EPR7577] (ab131263) at 1/5000 dilution (Purified)
Lanes 8-9 : Anti-IDH2 antibody [EPR7577] (ab131263) at 1/5000 dilution
Lane 1 : MOLT-4 (Human lymphoblastic leukemia T lymphoblast) whole cell lysate
Lane 2 : K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysate
Lane 3 : U-87 MG (Human glioblastoma-astrocytoma epithelial cell) whole cell lysate
Lane 4 : HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate
Lane 5 : Mouse liver lysate
Lane 6 : Rat liver lysate
Lane 7 : Mouse kidney lysate
Lane 8 : Rat kidney lysate
Lane 9 : Rat stomach lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 50 kDa -
This data was developed using ab230796, the same antibody clone in a different buffer formulation.
Flow Cytometry analysis of MCF7 (Human breast adenocarcinoma epithelial cell) cells labelling IDH2 with Purified ab230796 at 1:20 dilution (10 µg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150081) secondary antibody was used at 1:2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabelled control - Cell without incubation with primary antibody and secondary antibody (Blue). -
This data was developed using ab230796, the same antibody clone in a different buffer formulation.
Immunocytochemistry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling IDH2 with Purified ab230796 at 1:1000 dilution (0.2 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control. -
This data was developed using ab131263, the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver tissue sections labeling IDH2 with Purified ab131263 at 1:100 (2.11 µg/ml). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) . Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control. -
This data was developed using ab131263, the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse spleen tissue sections labeling IDH2 with Purified ab131263 at 1:100 (2.11 µg/ml). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) . Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control. -
This data was developed using ab131263, the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat spleen tissue sections labeling IDH2 with Purified ab131263 at 1:100 (2.11 µg/ml). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) . Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control. -
This WB data was generated using the same anti-IDH2 antibody clone, EPR7577, in a different buffer formulation (cat# ab131263).
Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: IDH2 knockout HAP1 cell lysate (20 µg)
Lane 3: K562 cell lysate (20 µg)
Lane 4: HepG2 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab131263 observed at 48 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab131263 was shown to recognize IDH2 when IDH2 knockout samples were used, along with additional cross-reactive bands. Wild-type and IDH2 knockout samples were subjected to SDS-PAGE. ab131263 and ab8245 (loading control to GAPDH) were both diluted 1/10 000 and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
-
Datasheet download
Certificate of Compliance
References (1)
ab230796 has been referenced in 1 publication.
- Gamradt S et al. Reduced mitochondrial respiration in T cells of patients with major depressive disorder. iScience 24:103312 (2021). PubMed: 34765928