Anti-IDH1 antibody (ab94571)
Key features and details
- Rabbit polyclonal to IDH1
- Suitable for: WB, IHC-P, ICC/IF
- Knockout validated
- Reacts with: Human
- Isotype: IgG
Overview
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Product name
Anti-IDH1 antibody
See all IDH1 primary antibodies -
Description
Rabbit polyclonal to IDH1 -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species ICC/IF HumanIHC-P HumanWB Human
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Immunogen
Synthetic peptide corresponding to Human IDH1 aa 350 to the C-terminus conjugated to keyhole limpet haemocyanin.
(Peptide available asab105207) -
Positive control
- This antibody gave a positive signal in HepG2 whole cell lysate as well as the following tissue lysates: Human kidney; Human testis. This antibody gave a positive result in IHC in the following FFPE tissue: Human normal liver.
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General notes
This product was previously labelled as Isocitrate dehydrogenase
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 or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration
Concentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab94571 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
GuaranteedTested applications are guaranteed to work and covered by our Abpromise guarantee.
PredictedPredicted to work for this combination of applications and species but not guaranteed.
IncompatibleDoes not work for this combination of applications and species.
Application Species ICC/IF HumanIHC-P HumanWB HumanAll applications MouseRatSheepCowApplication Abreviews Notes WB (2) Use a concentration of 1 µg/ml. Detects a band of approximately 47 kDa (predicted molecular weight: 47 kDa).IHC-P Use a concentration of 5 µg/ml.ICC/IF Use a concentration of 5 µg/ml.Notes WB
Use a concentration of 1 µg/ml. Detects a band of approximately 47 kDa (predicted molecular weight: 47 kDa).IHC-P
Use a concentration of 5 µg/ml.ICC/IF
Use a concentration of 5 µg/ml.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: 281235 Cow
- Entrez Gene: 3417 Human
- Entrez Gene: 15926 Mouse
- Entrez Gene: 24479 Rat
- Entrez Gene: 443257 Sheep
- Omim: 147700 Human
- SwissProt: Q9XSG3 Cow
- SwissProt: O75874 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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Western blot - Anti-IDH1 antibody (ab94571)
Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: IDH1 knockout HAP1 whole cell lysate (20 µg)
Lane 3: HeLa whole cell lysate (20 µg)
Lane 4: HepG2 whole cell lysate (20 µg)Lanes 1 - 4: Merged signal (red and green). Green - ab94571 observed at 46 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab94571 was shown to specifically recognize IDH1 in wild-type HAP1 cells along with additional cross reactive bands. No band was observed when IDH1 knockout samples were examined. Wild-type and IDH1 knockout samples were subjected to SDS-PAGE. Ab94571 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1 ug/ml and 1/10,000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.
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Western blot - Anti-IDH1 antibody (ab94571)All lanes : Anti-IDH1 antibody (ab94571) at 1 µg/ml
Lane 1 : Human kidney tissue lysate - total protein (ab30203)
Lane 2 : Human testis tissue lysate - total protein (ab30257)
Lane 3 : HepG2 (Human hepatocellular liver carcinoma 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: 47 kDa
Observed band size: 47 kDa
Additional bands at: 28 kDa, 78 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 3 minutes -
Immunocytochemistry/ Immunofluorescence - Anti-IDH1 antibody (ab94571)ICC/IF image of ab94571 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 (ab94571, 5µg/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) HeLa cells at 5µg/ml. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IDH1 antibody (ab94571)IHC image of IDH1 staining in Human normal liver formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab94571, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
Protocols
Datasheets and documents
References (3)
ab94571 has been referenced in 3 publications.
- Lee WD et al. Spatial-fluxomics provides a subcellular-compartmentalized view of reductive glutamine metabolism in cancer cells. Nat Commun 10:1351 (2019). PubMed: 30903027
- Niittykoski M et al. Immunohistochemical Characterization and Sensitivity to Human Adenovirus Serotypes 3, 5, and 11p of New Cell Lines Derived from Human Diffuse Grade II to IV Gliomas. Transl Oncol 10:772-779 (2017). PubMed: 28797937
- Jiang L et al. Reductive carboxylation supports redox homeostasis during anchorage-independent growth. Nature 532:255-8 (2016). WB . PubMed: 27049945
Images
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Western blot - Anti-IDH1 antibody (ab94571)
Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: IDH1 knockout HAP1 whole cell lysate (20 µg)
Lane 3: HeLa whole cell lysate (20 µg)
Lane 4: HepG2 whole cell lysate (20 µg)Lanes 1 - 4: Merged signal (red and green). Green - ab94571 observed at 46 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab94571 was shown to specifically recognize IDH1 in wild-type HAP1 cells along with additional cross reactive bands. No band was observed when IDH1 knockout samples were examined. Wild-type and IDH1 knockout samples were subjected to SDS-PAGE. Ab94571 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1 ug/ml and 1/10,000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.
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Western blot - Anti-IDH1 antibody (ab94571)All lanes : Anti-IDH1 antibody (ab94571) at 1 µg/ml
Lane 1 : Human kidney tissue lysate - total protein (ab30203)
Lane 2 : Human testis tissue lysate - total protein (ab30257)
Lane 3 : HepG2 (Human hepatocellular liver carcinoma 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: 47 kDa
Observed band size: 47 kDa
Additional bands at: 28 kDa, 78 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 3 minutes
-
Immunocytochemistry/ Immunofluorescence - Anti-IDH1 antibody (ab94571)ICC/IF image of ab94571 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 (ab94571, 5µg/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) HeLa cells at 5µg/ml.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IDH1 antibody (ab94571)
IHC image of IDH1 staining in Human normal liver formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab94571, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
