Description

  • Product name

    Recombinant Human HUS1 protein
    See all HUS1 proteins and peptides
  • Purity

    > 95 % SDS-PAGE.
    ab101944 is purified using conventional chromatography techniques.
  • Expression system

    Escherichia coli
  • Accession

  • Protein length

    Full length protein
  • Animal free

    No
  • Nature

    Recombinant
    • Species

      Human
    • Sequence

      MGSSHHHHHHSSGLVPRGSHMKFRAKIVDGACLNHFTRISNMIAKLAKTC TLRISPDKLNFILCDKLANGGVSMWCELEQENFFNEFQMEGVSAENNEIY LELTSENLSRALKTAQNARALKIKLTNKHFPCLTVSVELLSMSSSSRIVT HDIPIKVIPRKLWKDLQEPVVPDPDVSIYLPVLKTMKSVVEKMKNISNHL VIEANLDGELNLKIETELVCVTTHFKDLGNPPLASESTHEDRNVEHMAEV HIDIRKLLQFLAGQQVNPTKALCNIVNNKMVHFDLLHEDVSLQYFIPALS
    • Predicted molecular weight

      34 kDa including tags
    • Amino acids

      1 to 280
    • Tags

      His tag N-Terminus

Specifications

Our Abpromise guarantee covers the use of ab101944 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

    Mass Spectrometry

  • Mass spectrometry

    MALDI-TOF
  • Form

    Liquid
  • Concentration information loading...

Preparation and Storage

  • Stability and Storage

    Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.

    pH: 8.00
    Constituents: 0.316% Tris HCl, 40% Glycerol, 0.58% Sodium chloride

General Info

  • Alternative names

    • Checkpoint protein HUS1
    • hHUS1
    • Hus1
    • HUS1 (S. pombe) checkpoint homolog
    • HUS1 checkpoint homolog (S. pombe)
    • HUS1 Checkpoint Protein
    • HUS1 protein
    • HUS1+ - like protein
    • HUS1_HUMAN
    • Hydroxyurea-sensitive 1, S. pombe, homolog of
    see all
  • Function

    Component of the 9-1-1 cell-cycle checkpoint response complex that plays a major role in DNA repair. The 9-1-1 complex is recruited to DNA lesion upon damage by the RAD17-replication factor C (RFC) clamp loader complex. Acts then as a sliding clamp platform on DNA for several proteins involved in long-patch base excision repair (LP-BER). The 9-1-1 complex stimulates DNA polymerase beta (POLB) activity by increasing its affinity for the 3'-OH end of the primer-template and stabilizes POLB to those sites where LP-BER proceeds; endonuclease FEN1 cleavage activity on substrates with double, nick, or gap flaps of distinct sequences and lengths; and DNA ligase I (LIG1) on long-patch base excision repair substrates.
  • Tissue specificity

    Ubiquitous.
  • Sequence similarities

    Belongs to the HUS1 family.
  • Cellular localization

    Nucleus. Cytoplasm. In discrete nuclear foci upon DNA damage. According to PubMed:14500360, localized also in the cytoplasm. DNA damage induces its nuclear translocation. Shuttles between the nucleus and the cytoplasm.
  • Information by UniProt

Images

  • 15% SDS-PAGE analysis of 3µg ab101944.

References

ab101944 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-4 of 4 Abreviews or Q&A

Answer

Thank you for contacting us and I am sorry for the delay in getting back to you.

As mentioned in my previous email I have contacted the lab in regards to the HUS1 protein, ab101944 in order to try to understand why the results you have observed are not as expected. As I mentioned, the lot number you have been working was re-tested on an SDS-PAGE gel following the concerns raised by your colleague. I have reattached the results for your information. As can be seen from this image, I saw no concern in offering you a replacement from this lot to try again. A strong clean band of just under 40 kDa was observed with 3 ug of protein loaded.

We were just wondering if you could confirm a few further details in order to understand the problem further:

1. How many µL in total from the stock HUS1 and RAD1 protein were loaded on the gel?

2. How was the stain used with the gel? How long was the gel incubated in the stain?

3. Do you have access to a nanodrop where the protein content could be measured?

4. How were both the RAD1 and HUS1 proteins stored on receipt of the delivey?

I am sorry for the inconvinience casued by this problem. I look forward to recieving your reply.

Read More

Question
Answer

Thank you for your reply and for sharing that extra information.

From the details provided by the customer I cannot explain why they are not able to see the protein by SDS-PAGE when staining with the Coomassie stain. If they would like, I can provide a replacement vial of the protein. Or if they would prefer a credit note to be issued.

Thank you for your help in this case. I look forward to receiving your reply.

Read More

Question
Answer

I am sorry for the delay in getting back to you. I now have further information to share with you from the lab in regards to the HUS1 protein (ab101944).

Following the concerns raised by you, the protein has been re-run on an SDS-PAGE gel to observe the quality of the product. The results obtained can be viewed in the attachment of this email. As you can see, we have observed a strong clean band at the expected size (˜40 kDa) by Coomassie blue staining. This was performed with only 3 ug of protein loaded with the protein from the same batch you have used (GR100703-1).

In order to understand why you have not been observing the same, could you confirm a few further details:

1. When loading onto the SDS PAGE gel, you have said you load 0,25ng/ul to 1,5 ug/ul. Could you share an image of this gel and label each well with the total amount of protein in each well (i.e. if 3 ul of a 0,25ng/ul solution was diluted 1:1 with loading buffer you would be loading a total of 0.75 ng of protein in the well). If loading only in the ng range there may be difficulty in seeing the protein by Coomassie blue staining.

2. Could you explain how the Coomassie blue staining was performed? Was a protein ladder used at the same time, did this stain as expected?

3. You mention that you have stored the protein correctly. How has this been performed? Was the protein diluted prior to storage? What temperature was the protein stored at until use?

With this further information we may be able to ascertain why you have not been able to visualize the protein and hopefully resolve the problem.

I look forward to receiving your reply.

Read More

Answer

Thank you for contacting us.

I am sorry to hear you are experiencing difficulties with one of our products. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.

In order to understand what may be contributing to the problems encountered I would like to ask a few further questions if you would not mind:

1. How was the protein prepared prior to loading it onto the SDS-PAGE gel? Was a loading buffer added? Any reducing reagents? Was the protein boiled?

2. How much protein in total was loaded on the gel?

3. Which antibodies (anti Histidine –tag and anti-HUS1) were used? Catalogue number and manufacturer would be useful.

This information will allow us to understand the problems which may be contributing to the problems encountered and also help to investigate this case internally and initiate additional testing where necessary.

If the product was purchased in the last six months and is being used according to our Abpromise, we would be happy to replace or refund the antibody.

I look forward to receiving your reply.

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