Recombinant Human HINT1 protein (ab87362)

Reviews (1)Q&A (3)References (1)
SDS-PAGE - Recombinant Human HINT1 protein (ab87362)

    Key features and details

    • Expression system: Escherichia coli
    • Purity: > 90% SDS-PAGE
    • Suitable for: SDS-PAGE

    Description

    • Product name

      Recombinant Human HINT1 protein
      See all HINT1 proteins and peptides

    • Purity

      > 90 % SDS-PAGE.
      Purified by using conventional chromatography.

    • Expression system

      Escherichia coli

    • Protein length

      Full length protein

    • Animal free

      No

    • Nature

      Recombinant

      • Species

        Human

      • Sequence

        MADEIAKAQV ARPGGDTIFG KIIRKEIPAK IIFEDDRCLA FHDISPQAPT HFLVIPKKHI SQISVAEDDD ESLLGHLMIV GKKCAADLGL NKGYRMVVNE GSDGGQSVYH VHLHVLGGRQ MHWPPG

    Specifications

    Our Abpromise guarantee covers the use of ab87362 in the following tested applications.

    The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

    • Applications

      SDS-PAGE

    • Form

      Liquid
    • Concentration information loading...

    Preparation and Storage

    • Stability and Storage

      Shipped at 4°C. Store at +4°C.

      pH: 8.00
      Constituents: 0.316% Tris HCl, 10% Glycerol (glycerin, glycerine)

    General Info

    • Alternative names

      • Adenosine 5' monophosphoramidase
      • Adenosine 5''-monophosphoramidase
      • HINT 1
      • HINT1
      • HINT1_HUMAN
      • Histidine triad nucleotide binding protein 1
      • Histidine triad nucleotide-binding protein 1
      • PKCI 1
      • PKCI-1
      • PKCI1
      • PRKCNH1
      • Protein kinase C inhibitor 1
      • Protein kinase C interacting protein 1
      • Protein kinase C-interacting protein 1
      see all

    • Function

      Hydrolyzes adenosine 5'-monophosphoramidate substrates such as AMP-morpholidate, AMP-N-alanine methyl ester, AMP-alpha-acetyl lysine methyl ester and AMP-NH2.

    • Tissue specificity

      Widely expressed.

    • Sequence similarities

      Belongs to the HINT family.
      Contains 1 HIT domain.

    • Domain

      The histidine triad, also called HIT motif, forms part of the binding loop for the alpha-phosphate of purine mononucleotide.

    • Cellular localization

      Cytoplasm. Nucleus. Interaction with CDK7 leads to a more nuclear localization.
    • Information by UniProt

    Images

    • SDS-PAGE - Recombinant Human HINT1 protein (ab87362)
      SDS-PAGE - Recombinant Human HINT1 protein (ab87362)
      ab87362 on 15% SDS-PAGE (3µg)

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    References (1)

    Publishing research using ab87362? Please let us know so that we can cite the reference in this datasheet.

    ab87362 has been referenced in 1 publication.

    • Cortés-Montero E  et al. The Axonal Motor Neuropathy-Related HINT1 Protein Is a Zinc- and Calmodulin-Regulated Cysteine SUMO Protease. Antioxid Redox Signal 31:503-520 (2019). PubMed: 31088288

    Customer reviews and Q&As

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    1-4 of 4 Abreviews or Q&A

    Other (Hydrolyzes adenosine 5'-phosphoramidate) Abreview for HINT1 protein (Human)

     Good
    Abreviews
    abreview image
    Application
    Other - Do not use

    Review text: We have tested enzymatic activity of the HINT1 protein (ab87362)
    This enzyme hydrolyzes adenosine 5'-phosphoramidate (H2NPrA) to adenosine 5'-phosphate (rAMP).
    The protocol which we used in the assay.

    Fig1.
    Enzymatic reaction was carried out under the following conditions:
    Solution of substrate (adenosine 5'-phosphoramidate) at 1 mM were prepared in 20 mM Na-HEPES (pH 7.2) containing 0.5 mM of MgCl2 and the HINT1 (ab87362, 0.5, 1.0 or 2.0 µg) in 25 µl total volume.
    Through 15, 30, 60, 120 min and 18 h aliquots were taken (1 µl each). Aliquots were analyzed on TLC-plates in Silica gel 60 F254 (Merck) in system dioxan-isopropanol-25% aqueous ammonia-water – 8:7:3:2.
    As can be seen from the figure, we can assume:
    The hydrolysis of 50% amount of H2NPrA was 5 min with 2 µg of HINT1 2 mg/ml for 30 min with 1 µg of HINT1 and more than 60 min with 0.5 µg of HINT1.

    Fig2.
    Enzymatic reaction was carried out under the following conditions:
    Solution of substrate (adenosine 5'-phosphoramidate) at 1 mM were prepared in 20 mM Na-HEPES (pH 7.2) containing 0, 0.2 or 0.5 mM of MgCl2 and the HINT1 (ab87362, 0.5 or 1.0 µg) in 25 µl total volume.
    Through 15, 30, 60, 120 and 180 min aliquots were taken (1 µl each). Aliquots were analyzed on TLC-plates in Silica gel 60 F254 (Merck) in system dioxan-isopropanol-25% aqueous ammonia-water – 8:7:3:2.
    As can be seen from the figure, we can assume:
    HINT1 - magnesium-independent enzyme.
    The hydrolysis of 50% amount of H2NPrA was for 15-30 min with 1 µg of HINT1 and less than 120 min with 0.5 µg HINT1.

    When using HINT1 (ab87362) substrate hydrolysis was observed only up to adenosine 5’-monophosphate. During the enzymatic hydrolysis was not observed the formation of adenosine. In the absence of the enzyme in the reaction conditions adenosine 5'-phosphoramidate was stable.
    Sample: Human Recombinant protein

    Primary antibody (in addition to 'HINT1 protein (Human)')
    Primary antibody: None used

    Secondary antibody
    Secondary antibody: None used

    Mr. Alexander Shipitsyn

    Verified customer

    Submitted Nov 19 2012

    Question


    We agree to your proposal.
    We buy the HINT1 protein (Human) - ab87362 - throughwaiting for delivery.
    But we need to clarify about checking the activity of the enzyme.
    We plan to check HINT1 phosphoramidase activity for amide and
    monomorpholidate of adenosine monophosphate.
    We are likely to examine the enzyme in other phosphomonoamidates of
    various nucleosides.
    But check the phosphoramidase activity for amino acid derivatives of
    adenosine monophosphate is not in our plans.
    How and where should we send our Abreviev to take advantage of your
    offer?
    Sincerely,

    Read More
    Answer

    Thank you for your email.

    I am very pleased to hear you would like to accept our offer and test ab87362 for its biological acitivty. The code below will give you the equivalent price of ab87362 off any product in our catalogue.To redeem this offer, please submit an Abreview forfunctional studiesand include this code in the “Additional Comments” section so we know the Abreview is for this promotion. Please remember that submission of the Abreview is sufficient for the discount code to become active.

    DISCOUNT CODE: xxxxxxx
    Expiration date: 12th January 2012

    For more information on how to submit an Abreview, please visit the site:

    https://www.abcam.com/Abreviews

    Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways:

    1) Call to place your order and mention the code to our customer service department;
    2) Include the code in your fax order;
    3) Place your order on the web and enter the promotional code.

    Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

    The terms and conditions applicable to this offer can be found here:

    https://www.abcam.com/collaborationdiscount

    Read More

    Question

    Dear xxxx
    Yes, I undestend, that HINT1 protein (Human) (ab87362) is the full
    length recombinant human protein expressed in E. coli. The protein
    will therefore not contain any remnants from synthetic peptide
    synthesis.
    Perhaps I was not correct question. I would like to know whether the
    HINT1 protein stayed its ability to hydrolyze phosphoamide bond in
    adenosine 5'-monophosphoramidate substrates leads to the formation of
    adenosine 5'-monophosphosphate.
    Sincerely,

    Read More
    Answer

    Sorry for the delay in getting back to you.I now have some further information to share with you.

    Regrettably, theHINT1 protein (ab87362)biological activity has as yet not beentested. We do not therefore know if it has retained the enzymatic activity.I should mention thatthe proteinhasn’t been exposed to denaturing agents, so it is possible that it’s bioactive. However, as we have not tested it we would not be able to guarantee this.

    However, if you would like to purchase ab87362 and use it for its biological activity we can offer you a testing discount. This offer means that if you purchase ab87362 as normal, test the protein and let us know of the results through an Abreview (no matter whether positive or negative) you would be eligible for theequivalent price of ab87362 off any product in our catalogue. More information on this offer can be found here:

    www.abcam.com/collaborationdiscount

    Please note that theproduct to be tested must be purchased, tested, the Abreview submitted and the free product claimed within a 4 month period.

    If you would be interested in participating in this scheme please do let me know as a discount code needs to be issued prior to the purchase of ab87362.

    I hope this information has been of help. If you require any further information please do not hesitate to contact us again.

    Read More

    Question

    We want to buy HINT1 protein (Human) (ab87362).
    Could you tell us whether this protein exhibits phosphoramidaze
    activity?
    Sincerely,

    Read More
    Answer

    Thank you for contacting us and your interest in our products.

    I am sorry but I do not think I completely understand your question. The HINT1 protein (Human) (ab87362) is the full length recombinant human protein expressed in E. coli. The protein will therefore not contain any remnants from synthetic peptide synthesis.

    If this has not answered your question could you please explain the information you would like provided a little further?

    Otherwise, I wish you all the best with your research.

    Read More

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