Recombinant Hepatitis C Virus genotype 1a NS5 protein (ab49033)

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Overview

Description

  • Nature
    Recombinant
  • Source
    Escherichia coli
  • Amino Acid Sequence
    • Amino acids
      2212 to 2313

Specifications

Our Abpromise guarantee covers the use of ab49033 in the following tested applications.

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

  • Applications

    ELISA

    Western blot

    Flow Cytometry

  • Purity
    > 95 % SDS-PAGE.

  • Form
    Liquid
  • Concentration information loading...

Preparation and Storage

  • Stability and Storage

    Shipped at 4°C. After reconstitution store at -20ºC. Avoid freeze / thaw cycles.

    pH: 8.00
    Constituents: 0.2% Triton-X-100, 9% Urea, 0.395% Tris HCl, 50% Glycerol

General Info

  • Alternative names
    • HCV 1a NS5
    • Hepatitis C virus NS5A
    • Non-structural protein 5A
    • p56
  • Relevance
    Hepatitis C Virus is a positive, single stranded RNA virus in the Flaviviridae family. The genome is approximately 10,000 nucleotides and encodes a single polyprotein of about 3,000 amino acids. The polyprotein is processed by host cell and viral proteases into three major structural proteins and several non structural proteins necessary for viral replication. Several different genotypes of HCV with slightly different genomic sequences have since been identified that correlate with differences in response to treatment with interferon alpha. NS5A is a ~56 kDa pleiotropic protein with key roles in both viral RNA replication and modulation of the physiology of the host cell. It's exact role is not currently known (2008). NS5B (non-structural protein 5B) is an RNA-dependant RNA polymerase responsible for replication of the hepatitis C viral genome, and is currently a principal target for chemotherapeutic inhibition of HCV replication. Hepatitis C virus (HCV) can cause chronic hepatitis, cirrhosis and hepatocellular carcinoma. At present there is no vaccine effective against HCV.
  • Cellular localization
    Endoplasmic reticulum membrane

References

ab49033 has not yet been referenced specifically in any publications.

Publishing research using ab49033? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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Question


I’ve attached the protocol I used for the Western blot. I’m trying to find a good antibody to HCV NS5A so that I can select stable clones overexpressing the protein. So far, I haven’t been able to pick up any NS5A using two proteins from Abcam, ab49033 and ab68096, and high concentrations (1:100, 1:250) of four different antibodies (ab20342, sc-57776, ab20773, ab2594).

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Answer

Thanks for sending along your protocol.

1. ab49033 is unlikely to be detected by these antibodies as it only contains about 100 aa of the total protein. As described on datasheet this product containsamino acids"corresponding to amino acids 2212-2313 of Hepatitis C Virus NS5" The epitopes recognized by these antibodies would have to recognize NS5A within this small region to work.

2. Usingponceau S stain post transfer, have you been able to determine protein is present on membrane after transfer?

3. Regarding your protocol, a) Boiling of samplesis not recommended; instead heat at 70 deg C for 10min prior to loading gel b) Try blocking with 5% BSA instead of milk c) Use 1:1000 for primary antibodies and 1:5000 for secondary antibodies.

I look forward to hearing if these suggestions prove useful.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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