- Datasheet
- Specific References
- Protocols
Overview
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Product nameAnti-IDH2 antibody
See all IDH2 primary antibodies -
DescriptionRabbit polyclonal to IDH2
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Host speciesRabbit
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Tested applicationsSuitable for: WB, ICC/IFmore details
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Species reactivityReacts with: Human
Predicted to work with: Mouse, Cow, Pig
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Immunogen
Synthetic peptide conjugated to KLH derived from within residues 150 - 250 of Human IDH2.
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Positive control
- This antibody gave a positive signal in Human Heart, Skeletal Muscle, Liver and Brain Tissue Lysates, and in HepG2, DU 145 and SHSY5Y whole cell lysates.
Properties
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FormLiquid
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Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
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Storage bufferPreservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4 -
Concentration information loading... -
PurityImmunogen affinity purified
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ClonalityPolyclonal
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IsotypeIgG
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Research areas
Associated products
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Compatible Secondaries
Applications
Our Abpromise guarantee covers the use of ab94359 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 |
|---|---|---|
| WB | Use a concentration of 1 µg/ml. Detects a band of approximately 47 kDa (predicted molecular weight: 51 kDa). | |
| ICC/IF | Use a concentration of 1 µg/ml. |
Target
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FunctionPlays a role in intermediary metabolism and energy production. It may tightly associate or interact with the pyruvate dehydrogenase complex.
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Involvement in diseaseD-2-hydroxyglutaric aciduria 2
Glioma
enetic variations are associated with cartilaginous tumors such as enchondroma or chondrosarcoma. -
Sequence similaritiesBelongs to the isocitrate and isopropylmalate dehydrogenases family.
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Post-translational
modificationsAcetylation at Lys-413 dramatically reduces catalytic activity. Deacetylated by SIRT3. -
Cellular localizationMitochondrion.
- Information by UniProt
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Database links
- Entrez Gene: 327669 Cow
- Entrez Gene: 3418 Human
- Entrez Gene: 269951 Mouse
- Entrez Gene: 397603 Pig
- Omim: 147650 Human
- SwissProt: Q04467 Cow
- SwissProt: P48735 Human
- SwissProt: P54071 Mouse
see all -
Alternative names
- D2HGA2 antibody
- ICD-M antibody
- IDH antibody
see all
Images
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Western blot - Anti-IDH2 antibody (ab94359)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 - ab94359 observed at 47 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab94359 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. ab94359 at a concentration of 1ug/mL and ab8245 (loading control to GAPDH) diluted to 1/2000 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. -
Western blot - Anti-IDH2 antibody (ab94359)All lanes : Anti-IDH2 antibody (ab94359) at 1 µg/ml
Lane 1 : Human heart tissue lysate - total protein (ab29431)
Lane 2 : Human skeletal muscle tissue lysate - total protein (ab29330)
Lane 3 : Human liver tissue lysate - total protein (ab29889)
Lane 4 : Human brain tissue lysate - total protein (ab29466)
Lane 5 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
Lane 6 : DU 145 (Human prostate carcinoma cell line) Whole Cell Lysate
Lane 7 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 51 kDa
Observed band size: 47 kDa (why is the actual band size different from the predicted?)
Additional bands at: 35 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 30 seconds
The predicted molecular weight of the IDH2 protein is 51-kDa. However, the protein sequence contains a 39 residue transit peptide at the N-terminus. This might explain the migration of the protein at a lower molecular weight than expected. -
Immunocytochemistry/ Immunofluorescence - Anti-IDH2 antibody (ab94359)ICC/IF image of ab94359 stained HepG2 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab94359, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 100% methanol fixed (5 min) MCF7 cells at 1µg/ml, and in 4% PFA fixed (10 min) HepG2 cells at 1µg/ml.
References
ab94359 has not yet been referenced specifically in any publications.
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