Recombinant Anti-IDH2 antibody [EPR7576] (ab129180)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR7576] to IDH2
- Suitable for: Flow Cyt (Intra), WB, IHC-P
- Knockout validated
- Reacts with: Mouse, Human
Related conjugates and formulations
Overview
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Product name
Anti-IDH2 antibody [EPR7576]
See all IDH2 primary antibodies -
Description
Rabbit monoclonal [EPR7576] to IDH2 -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, IHC-Pmore details
Unsuitable for: ICC/IF or IP -
Species reactivity
Reacts with: Mouse, Human
Predicted to work with: Rat -
Immunogen
Synthetic peptide within Human IDH2 aa 50-150. The exact sequence is proprietary.
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Positive control
- Human thyroid gland carcinoma tissue; Molt-4, K562, 293T and HepG2 whole cell lysate (ab7900).
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 9% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA, 50% Tissue culture supernatant -
Concentration information loading...
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Purity
Tissue culture supernatant -
Clonality
Monoclonal -
Clone number
EPR7576 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab129180 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 |
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Flow Cyt (Intra) |
1/1000 - 1/10000.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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WB | (3) |
1/1000 - 1/10000. Detects a band of approximately 45 kDa (predicted molecular weight: 51 kDa).
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IHC-P |
1/250 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Notes |
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Flow Cyt (Intra)
1/1000 - 1/10000. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
WB
1/1000 - 1/10000. Detects a band of approximately 45 kDa (predicted molecular weight: 51 kDa). |
IHC-P
1/250 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
Target
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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
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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
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All lanes : Anti-IDH2 antibody [EPR7576] (ab129180) at 1/1000 dilution
Lane 1 : Wild-type Jurkat cell lysate
Lane 2 : IDH2 knockout Jurkat cell lysate
Lane 3 : MOLT-4 cell lysate
Lane 4 : HEK-293 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 51 kDa
Observed band size: 48 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-IDH2 antibody [EPR7576] 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, ab129180 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.
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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 - ab129180 observed at 47 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab129180 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. ab129180 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/2000 respectively 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. -
ab129180, at 1/250 dilution, staining IDH2 in Formalin-fixed, Paraffin-embedded Human thyroid gland carcinoma by Immunohistochemistry.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Overlay histogram showing MCF7 cells stained with ab129180 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab129180, 1/10000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in MCF7 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
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All lanes : Anti-IDH2 antibody [EPR7576] (ab129180) at 1/1000 dilution
Lane 1 : Molt-4 cell lysate
Lane 2 : K562 cell lysate
Lane 3 : 293T cell lysate
Lane 4 : HepG2 cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-rabbit HRP
Predicted band size: 51 kDa
Observed band size: 45 kDa why is the actual band size different from the predicted?
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (5)
ab129180 has been referenced in 5 publications.
- Benlamara S et al. Distinctive Krebs cycle remodeling in iPSC-derived neural and mesenchymal stem cells. Biochem Biophys Res Commun 511:658-664 (2019). PubMed: 30826061
- Pichitpunpong C et al. Phenotypic subgrouping and multi-omics analyses reveal reduced diazepam-binding inhibitor (DBI) protein levels in autism spectrum disorder with severe language impairment. PLoS One 14:e0214198 (2019). PubMed: 30921354
- Tischler J et al. Metabolic regulation of pluripotency and germ cell fate through a-ketoglutarate. EMBO J 38:N/A (2019). PubMed: 30257965
- White K et al. Loss of IDH2 Accelerates Age-related Hearing Loss in Male Mice. Sci Rep 8:5039 (2018). PubMed: 29567975
- Gao J et al. Trilobatin Protects Against Oxidative Injury in Neuronal PC12 Cells Through Regulating Mitochondrial ROS Homeostasis Mediated by AMPK/Nrf2/Sirt3 Signaling Pathway. Front Mol Neurosci 11:267 (2018). PubMed: 30104959