Anti-IDH2 antibody [5F11] (ab55271)
Key features and details
- Mouse monoclonal [5F11] to IDH2
- Suitable for: IHC-P, Flow Cyt, ICC/IF, WB
- Knockout validated
- Reacts with: Human, Recombinant fragment
- Isotype: IgG1
Overview
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Product name
Anti-IDH2 antibody [5F11]
See all IDH2 primary antibodies -
Description
Mouse monoclonal [5F11] to IDH2 -
Host species
Mouse -
Tested applications
Suitable for: IHC-P, Flow Cyt, ICC/IF, WBmore details -
Species reactivity
Reacts with: Human, Recombinant fragment -
Immunogen
Recombinant fragment corresponding to Human IDH2 aa 354-451.
Sequence:HYREHQKGRPTSTNPIASIFAWTRGLEHRGKLDGNQDLIRFAQMLEKVCV ETVESGAMTKDLAGCIHGLSNVKLNEHFLNTTDFLDTIKSNLDRALGR
Database link: P48735 -
Positive control
- IHC-P: Human colon. Flow cyt: MCF7 cells. WB: Transfected 293T cells.
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General notes
This product was changed from ascites to tissue culture supernatant on 13th Feb 2019. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles. -
Storage buffer
pH: 7.4 -
Concentration information loading...
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Purity
Tissue culture supernatant -
Clonality
Monoclonal -
Clone number
5F11 -
Isotype
IgG1 -
Light chain type
kappa -
Research 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 ab55271 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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IHC-P |
Use at an assay dependent concentration.
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Flow Cyt |
Use at an assay dependent concentration.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
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ICC/IF |
Use at an assay dependent concentration.
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WB |
Use at an assay dependent concentration.
This antibody has only been tested in WB against the recombinant fragment used as immunogen. We have no data on the detection of endogenous protein. |
Notes |
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IHC-P
Use at an assay dependent concentration. |
Flow Cyt
Use at an assay dependent concentration. ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
ICC/IF
Use at an assay dependent concentration. |
WB
Use at an assay dependent concentration. This antibody has only been tested in WB against the recombinant fragment used as immunogen. We have no data on the detection of endogenous protein. |
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
- Omim: 147650 Human
- SwissProt: P48735 Human
- Unigene: 596461 Human
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Alternative names
- D2HGA2 antibody
- ICD-M antibody
- IDH antibody
see all
Images
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All lanes : Anti-IDH2 antibody [5F11] (ab55271) 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.
Observed band size: 48 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-IDH2 antibody [5F11] staining at 1/1000 dilution, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (ab181602) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab55271 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-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) at 1/20000 dilution.
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Western blot against tagged recombinant protein immunogen using ab55271 IDH2 antibody at 1ug/ml. Predicted band size of immunogen is 37 kDa
This image was generated using the ascites version of the product.
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IDH2 antibody (ab55271) used in immunohistochemistry at 3ug/ml on formalin fixed and paraffin embedded human colon.
This image was generated using the ascites version of the product.
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ICC/IF image of ab55271 stained Mcf7 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 (ab55271, 10µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse 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 image was generated using the ascites version of the product.
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All lanes : Anti-IDH2 antibody [5F11] (ab55271)
Lane 1 : IDH2 in transfected 293T cell line
Lane 2 : Non-transfected lysateThe blocking agent used is 5% milk.
This image was generated using the ascites version of the product.
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Overlay histogram showing MCF7 cells stained with ab55271 (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 (ab55271, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. 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.
This image was generated using the ascites version of the product.
Protocols
Datasheets and documents
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Datasheet download
References (39)
ab55271 has been referenced in 39 publications.
- Lin AP et al. MYC, mitochondrial metabolism and O-GlcNAcylation converge to modulate the activity and subcellular localization of DNA and RNA demethylases. Leukemia 36:1150-1159 (2022). PubMed: 34997181
- Han W et al. Targeting HOTAIRM1 ameliorates glioblastoma by disrupting mitochondrial oxidative phosphorylation and serine metabolism. iScience 25:104823 (2022). PubMed: 35992092
- Sabatier M et al. Activation of Vitamin D Receptor Pathway Enhances Differentiating Capacity in Acute Myeloid Leukemia with Isocitrate Dehydrogenase Mutations. Cancers (Basel) 13:N/A (2021). PubMed: 34680392
- Gambichler T et al. Altered hydroxymethylation in cutaneous squamous cell carcinoma and keratoacanthoma. Br J Dermatol N/A:N/A (2020). PubMed: 32407588
- Gambichler T et al. A study on DNA hydroxymethylation in Kaposi sarcoma and cutaneous angiosarcoma. J Eur Acad Dermatol Venereol N/A:N/A (2020). PubMed: 32279358