

Recombinant Anti-IDH1 antibody [EPR21002] (ab230949)
- Datasheet
- References
- Protocols
Overview
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Product name
Anti-IDH1 antibody [EPR21002]
See all IDH1 primary antibodies -
Description
Rabbit monoclonal [EPR21002] to IDH1 -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-P, Flow Cyt, IPmore details
Unsuitable for: ICC/IF -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide within Human IDH1 aa 100-200. The exact sequence is proprietary.
Database link: O75874 -
Positive control
- WB: SH-SY5Y, Raji, C2C12, Neuro-2a, HepG2, HeLa, C6 and PC-12 whole cell lysates; Human fetal brain and fetal kidney lysates. IHC-P: Human stomach and endometrium cancer tissues; Mouse and rat kidney tissues. Flow Cyt: HeLa cells. IP: SH-SY5Y whole cell lysate.
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General notes
This product was previously labelled as Isocitrate dehydrogenase
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.
This product is a recombinant rabbit monoclonal antibody.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR21002 -
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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Recombinant Protein
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Related Products
Applications
Our Abpromise guarantee covers the use of ab230949 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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WB | 1/1000. Detects a band of approximately 47 kDa (predicted molecular weight: 46 kDa). | |
IHC-P | 1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. | |
Flow Cyt | 1/50. | |
IP | 1/30. |
Target
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Involvement in disease
Glioma
Genetic variations are associated with cartilaginous tumors such as enchondroma or chondrosarcoma. Mutations of Arg-132 to Cys, Gly or His abolish the conversion of isocitrate to alpha-ketoglutarate. Instead, alpha-ketoglutarate is converted to R(-)-2-hydroxyglutarate. -
Sequence similarities
Belongs to the isocitrate and isopropylmalate dehydrogenases family. -
Post-translational
modificationsAcetylation at Lys-374 dramatically reduces catalytic activity. -
Cellular localization
Cytoplasm. Peroxisome. - Information by UniProt
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Database links
- Entrez Gene: 3417 Human
- Entrez Gene: 15926 Mouse
- Entrez Gene: 24479 Rat
- Omim: 147700 Human
- SwissProt: O75874 Human
- SwissProt: O88844 Mouse
- SwissProt: P41562 Rat
- Unigene: 593422 Human
see all -
Alternative names
- Cytosolic NADP isocitrate dehydrogenase antibody
- Cytosolic NADP-isocitrate dehydrogenase antibody
- Epididymis luminal protein 216 antibody
see all
Images
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All lanes : Anti-IDH1 antibody [EPR21002] (ab230949) at 1/1000 dilution
Lane 1 : Wild type HAP1 whole cell lysate
Lane 2 : IDH1 knockout HAP1 whole cell lysate
Lane 3 : SH-SY5Y (human neuroblastoma epithelial cell), whole cell lysate
Lysates/proteins at 10 µg per lane.
Predicted band size: 46 kDa
Exposure time: 62 secondsab230949 was shown to specifically react with IDH1 in wild-type HAP1 cells as signal was lost in IDH1 knockout cells. Wild-type and IDH1 knockout samples were subjected to SDS-PAGE. ab230949 and ab181602 (Rabbit anti-GAPDH loading control) were incubated 1 hour at room temperature at 1/1000 dilution and 1/200,000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) secondary antibody at 1/100,000 dilution for 1 hour at room temperature before imaging.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IDH1 antibody [EPR21002] (ab230949)
Immunohistochemical analysis of paraffin-embedded rat kidney tissue labeling IDH1 with ab230949 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic and nuclear staining in rat kidney (PMID:30153799). Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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All lanes : Anti-IDH1 antibody [EPR21002] (ab230949) at 1/1000 dilution
Lane 1 : SH-SY5Y (human neuroblastoma cell line from bone marrow) whole cell lysate at 20 µg
Lane 2 : Raji (human Burkitt's lymphoma cell line) whole cell lysate at 20 µg
Lane 3 : C2C12 (mouse myoblast cell line) whole cell lysate at 20 µg
Lane 4 : Neuro-2a (mouse neuroblastoma cell line) whole cell lysate at 20 µg
Lane 5 : Human fetal brain lysate at 20 µg
Lane 6 : Human fetal kidney lysate at 20 µg
Lane 7 : HepG2 (human liver hepatocellular carcinoma cell line) whole cell lysate at 20 µg
Lane 8 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 9 : C6 (rat glial tumor cell line) whole cell lysate at 10 µg
Lane 10 : PC-12 (rat adrenal gland pheochromocytoma cell line) whole cell lysate at 10 µg
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 46 kDa
Observed band size: 47 kDa why is the actual band size different from the predicted?Exposure times: Lanes 1-6: 3 minutes; Lanes 7-8: 15 seconds; Lanes 9-10: 58 seconds.
Blocking/Dilution buffer: 5% NFDM/TBST.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IDH1 antibody [EPR21002] (ab230949)
Immunohistochemical analysis of paraffin-embedded mouse kidney tissue labeling IDH1 with ab230949 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic and nuclear staining in mouse kidney (PMID:30153799). Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IDH1 antibody [EPR21002] (ab230949)
Immunohistochemical analysis of paraffin-embedded human endometrium cancer tissue labeling IDH1 with ab230949 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic and nuclear staining in human endometrium cancer (PMID:29921847). Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IDH1 antibody [EPR21002] (ab230949)
Immunohistochemical analysis of paraffin-embedded human stomach tissue labeling IDH1 with ab230949 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic and nucleus staining in human stomach (PMID:27466503). Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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IDH1 was immunoprecipitated from 0.35 mg of SH-SY5Y (human neuroblastoma cell line from bone marrow) whole cell lysate with ab230949 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab230949 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
Lane 1: SH-SY5Y whole cell lysate 10 µg (Input).
Lane 2: ab230949 IP in SH-SY5Y whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab230949 in SH-SY5Y whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 30 seconds.
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Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cell line labeling IDH1 with ab230949 at 1/60 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.
Datasheets and documents
References
ab230949 has not yet been referenced specifically in any publications.