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    idh2-antibody-ab55271.pdf

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Signal Transduction Metabolism Energy Metabolism
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Anti-IDH2 antibody (ab55271)

  • Datasheet
Submit a review Q&A (3)References (30)

Product price, shipping and contact information

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Western blot - Anti-IDH2 antibody (ab55271)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IDH2 antibody (ab55271)
  • Immunocytochemistry/ Immunofluorescence - Anti-IDH2 antibody (ab55271)
  • Western blot - Anti-IDH2 antibody (ab55271)
  • Flow Cytometry - Anti-IDH2 antibody (ab55271)

Key features and details

  • Mouse monoclonal to IDH2
  • Suitable for: IHC-P, Flow Cyt, ICC/IF, WB
  • Reacts with: Human, Recombinant fragment
  • Isotype: IgG1

You may also be interested in

Assay
Product image
Frataxin Protein Quantity Dipstick Assay Kit (ab109881)
Protein
Product image
Recombinant human IDH2 protein (ab198092)
Secondary
Product image
Goat Anti-Mouse IgG H&L (HRP) (ab205719)

View more associated products

Overview

  • Product name

    Anti-IDH2 antibody
    See all IDH2 primary antibodies
  • Description

    Mouse monoclonal to IDH2
  • Host species

    Mouse
  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    ICC/IF
    Human
    IHC-P
    Human
    WB
    Human
    Recombinant fragment
    See all applications and species data
  • Immunogen

    Recombinant fragment corresponding to Human IDH2 aa 354-451.
    Sequence:

    HYREHQKGRPTSTNPIASIFAWTRGLEHRGKLDGNQDLIRFAQMLEKVCV ETVESGAMTKDLAGCIHGLSNVKLNEHFLNTTDFLDTIKSNLDRALGR


    Database link: P48735
    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI
  • Positive control

    • IHC-P: Human colon. Flow cyt: MCF7 cells. WB: Transfected 293T cells.
  • 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.

Properties

  • 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...
  • Purity

    Tissue culture supernatant
  • Clonality

    Monoclonal
  • Isotype

    IgG1
  • Light chain type

    kappa
  • Research areas

    • Signal Transduction
    • Metabolism
    • Energy Metabolism
    • Signal Transduction
    • Metabolism
    • Mitochondrial
    • Cancer
    • Cancer Metabolism
    • Metabolic signaling pathway
    • Integration of energy metabolism
    • Metabolism
    • Pathways and Processes
    • Mitochondrial Metabolism
    • Mitochondrial markers
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Energy transfer pathways
    • Energy Metabolism
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Energy transfer pathways
    • Integration of energy
    • Metabolism
    • Types of disease
    • Cancer

Associated products

  • Compatible Secondaries

    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
    • Goat Anti-Mouse IgG H&L (HRP) (ab205719)
    • Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed (ab96879)
  • Isotype control

    • Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control (ab170190)
    • Mouse IgG1, Kappa Monoclonal [B11/6] - Isotype Control (ab91353)
  • Recombinant Protein

    • Recombinant human IDH2 protein (ab198092)

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.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
Flow Cyt
Human
ICC/IF
Human
IHC-P
Human
WB
Human
Recombinant fragment
Application Abreviews Notes
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.

Notes
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

  • 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
    modifications

    Acetylation at Lys-413 dramatically reduces catalytic activity. Deacetylated by SIRT3.
  • Cellular localization

    Mitochondrion.
  • Target information above from: UniProt accession P48735 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 3418 Human
    • Omim: 147650 Human
    • SwissProt: P48735 Human
    • Unigene: 596461 Human
    • Alternative names

      • D2HGA2 antibody
      • ICD-M antibody
      • IDH antibody
      • IDH2 antibody
      • IDHM antibody
      • IDHP_HUMAN antibody
      • IDP antibody
      • IDPM antibody
      • Isocitrate dehydrogenase [NADP], mitochondrial antibody
      • Isocitrate dehydrogenase 2 (NADP+), mitochondrial antibody
      • mNADP-IDH antibody
      • NADP(+)-specific ICDH antibody
      • Oxalosuccinate decarboxylase antibody
      see all

    Images

    • Western blot - Anti-IDH2 antibody (ab55271)
      Western blot - Anti-IDH2 antibody (ab55271)

      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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IDH2 antibody (ab55271)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IDH2 antibody (ab55271)

      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.

    • Immunocytochemistry/ Immunofluorescence - Anti-IDH2 antibody (ab55271)
      Immunocytochemistry/ Immunofluorescence - Anti-IDH2 antibody (ab55271)

      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.

    • Western blot - Anti-IDH2 antibody (ab55271)
      Western blot - Anti-IDH2 antibody (ab55271)
      All lanes : Anti-IDH2 antibody (ab55271)

      Lane 1 : IDH2 in transfected 293T cell line
      Lane 2 : Non-transfected lysate


      The blocking agent used is 5% milk.

      This image was generated using the ascites version of the product.

    • Flow Cytometry - Anti-IDH2 antibody (ab55271)
      Flow Cytometry - Anti-IDH2 antibody (ab55271)

      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

    • Flow cytometry protocols
    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet
  • References (30)

    Publishing research using ab55271? Please let us know so that we can cite the reference in this datasheet.

    ab55271 has been referenced in 30 publications.

    • 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
    • Shi F  et al. Wild-type IDH2 contributes to Epstein-Barr virus-dependent metabolic alterations and tumorigenesis. Mol Metab 36:100966 (2020). PubMed: 32224436
    • Aljohani AI  et al. The prognostic significance of wild-type isocitrate dehydrogenase 2 (IDH2) in breast cancer. Breast Cancer Res Treat 179:79-90 (2020). PubMed: 31599393
    • Zhang Y  et al. IDH2 compensates for IDH1 mutation to maintain cell survival under hypoxic conditions in IDH1-mutant tumor cells. Mol Med Rep 20:1893-1900 (2019). PubMed: 31257503
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-3 of 3 Abreviews or Q&A

    Question

    In continuation to our previous correspondence, the customer tried your suggestions (she also diluted the antibody 1/1000 and usedNitrocellulose membrane). Unfortunately, these did not improve the results. The customer also performed WB with IDH2 antibody from Human Atlas (Sigma) and this showed the predicted product stronger than non specific, whereas Abcam's ab showed strong unspecific band and very weak or invisible specific bands. Please find the comparative gel image.

    Read More

    Abcam community

    Verified customer

    Asked on May 04 2012

    Answer

    Thank you for your message and for providing this further information.
    I am sorry to hear the suggestions made have not improved the results on this occasion. I appreciate the time your customer has spent on these experiments, and would like to offer a free of charge replacement or credit note in compensation (providing the product has been purchased in the last 180 days). In order to arrange this, I would appreciate if you could confirm the order number and date of purchase?
    Thank you for your continued cooperation. I look forward to hearing from you with details of how your customer would like to proceed.

    Read More

    Abcam Scientific Support

    Answered on May 04 2012

    Question

    Dear Tech Support Team,
    Please find below the details of customer's complaint. Attached is the image of the gel.
    Please advise.

    Read More

    Abcam community

    Verified customer

    Asked on Mar 13 2012

    Answer

    Thank you for taking the time to complete our questionnaire and contact us. I am sorry to hear the customer has had difficulty obtaining satisfactory results from this IDH2 antibody.

    The details you have kindly provided will enable us to investigate this case for you and also gives us vital information for our monitoring of product quality.

    I appreciate the time spent in the laboratory and understand the concerns. It is regrettable the results have not been successful. However, a band around 51 kDa seems to be present (as expected from the Isocitrate dehydrogenase according to SwissProt ID P48735), and therefore I would like to offer some suggestions to help optimise the results:

    1) For mitochondrial proteins such as IDH2 a specific lysis buffer (RIPA buffer) or even fractionation of cell compartments may help to optimise the results. Please find details attached in our guide.

    2) Loading less sample may help to reduce the background: Usually, 20 -30 ug protein per lane is sufficient.

    3) The choice of membrane may give high background: Nitrocellulose membrane is considered to give less background than PVDF, so I would recommend trying it with nitrocellulose membranes.

    4) When testing our antibodies, our lab uses 5% BSA as a blocking reagent, so I recommend switching to this instead of milk. For unknown reasons some antibodies give stronger, more specific signals on blots blocked with BSA instead of milk, so doing this may improve the results you are seeing, and reduce the non-specific bands. Also, use one blocking reagent throughout the protocol, i.e. if blocked with BSA then also use it in the antibody dilution buffers.

    4) You might reduce the concentration of the primary antibody even further (to 1 ug/ml or even less).

    5) The secondary antibody may be binding non-specifically or reacting with the blocking reagent. I would suggest to run a secondary control without primary antibody to assess this issue.

    6) If possible, you could run the gel a bit longer to better separate the higher molecular weight proteins. You then might be able to distinguish different post-translationally modified IDH2 forms.

    We are happy to offer this technical support. If these suggestions have not yet been tried, I would appreciate if you can consider trying the suggestions. I hope you would agree that it would be preferable if we can get the experiment working and giving good results.

    I hope this information is helpful, thank you for your cooperation. Should the suggestions not improve the results, please do not hesitate to contact me again with the further requested details.

    Read More

    Abcam Scientific Support

    Answered on Mar 13 2012

    Question

    Bonjour, J'ai commandé un anticorps (anti IDH2, ref ab55271) mais rien n'est inscrit sur la fiche technique quant au blocage des sites non spé (avec serum bovine albumine ou avec du lait ? et à quel pourcentage ? ) Merci pour votre réponse Cordialement  

    Read More

    Abcam community

    Verified customer

    Asked on Nov 16 2011

    Answer

    Merci de nous avoir contactés. L'anticorps anti-IDH2 ab55271 a récemment été testé sur un lysat de cellules tranfectées. Une bande de 51 kDa, correspondant à la IDH2, a été observée. L'agent bloquant utilisé est le lait à 5%. La fiche technique de l'anti-IDH2 ab55271 sera mise à jour très prochainement. J'espère que ces informations vous seront utiles. N'hésitez pas à nous contacter de nouveau si vous avez d'autres questions.

    Read More

    Abcam Scientific Support

    Answered on Nov 16 2011

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