Recombinant Human HINT1 protein (ab87362)

Reviews (1)Q&A (3)

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

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)

References

ab87362 has not yet been referenced specifically in any publications.

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

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)

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Application
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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,

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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

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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.

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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.

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