Overview

Description

  • Nature
    Recombinant
  • Source
    Escherichia coli
  • Amino Acid Sequence
    • Amino acids
      1658 to 1863
  • Conjugation
    FITC. Ex: 493nm, Em: 528nm

Specifications

Our Abpromise guarantee covers the use of ab49029 in the following tested applications.

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

  • Biological activity
    Reacts strongly with human HCV positive serum.
  • Applications

    Western blot

    ELISA

    SDS-PAGE

  • Purity
    > 95 % SDS-PAGE.

  • Form
    Liquid
  • Additional notes
    Reacts strongly with human HCV positive serum.
  • Concentration information loading...

Preparation and Storage

  • Stability and Storage

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.

    pH: 8.00
    Constituents: 48% Urea, 0.078% Beta mercaptoethanol, 0.316% Tris HCl

General Info

  • Alternative names
    • HCV
    • HCV NS4
    • Hepatitis C Virus nonstructural antigen 4
    • Non structural protein 4A
    • Non structural protein 4B
    • NS4A
    • NS4B
    • p27
    • p8
    see all
  • Relevance
    NS4 A/B are two of the seven nonstructural (NS) proteins making up the HCV polyprotein. The NS proteins recruit the viral genome into an RNA replication complex, which is associated with rearranged cytoplasmic membranes. NS4A acts as a cofactor with the NS3 serine protease and stabilizes its folding. The NS3-NS4A complex is essential for the activation of the latter and allows membrane anchorage of NS3. HCV 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. HCV is responsible for a large proportion of worldwide chronic viral hepatitides. Most of these infections develop into chronic hepatitis, which often progresses to liver cirrhosis and hepatocellular carcinoma. At present, (unlike hepatitis A and B), there is no vaccine to prevent hepatitis C infection. The hepatitis C virus (HCV) nonstructural protein 4B (NS4B) is a relatively hydrophobic 27-kDa protein. The 4A protein has a molecular weight of 6 kDa.
  • Cellular localization
    Endoplasmic reticulum; multi-pass membrane protein.

References

ab49029 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Question
Answer


The ELISA protocol is below.
If antigen is diluted at least 1 to 10 the 8M Urea will not be a problem.


ELISA protocol

1. Dilute HCV antigen to 0.1 mg/25 ml (4.0 mg/ml) in coating buffer (0.01M Na2CO3; 0.01M Na3EDTA; 0.014 M b-mercaptoethanol) e.g. for 10 wells requiring 10 X 25 ml = 250 ml use 10 ml of stock antigen into coating buffer.

2. Coat each well with 25 ml (0.1 mg) of antigen in coating buffer by adding to wells of a pretreated ELISA plate, covering with plastic film and incubating at 4oC overnight. This may be performed at 37oC for 1 hour.

3. Shake coating solution out of wells and wash 2 X with water.

4. Block wells by adding 50 ml 3% BSA/PBS per well and incubate for 1 hour at 37oC, wrapped in plastic or in a moist sealed container.

5. Shake off blocking solution (without washing or drying) and add primary antibody preparation diluted in 1% BSA/PBS, 25 ml per well. The recommended antibody dilution should be: 1/10, 1/100, 1/1000, 1/10000, and undiluted.

6. Wrap and incubate at 37oC for 1 hour.

7. Shake off the antibody solution and wash 10 X with PBS/Tween-20 (0.05%) or distilled water.

8. Add antibody enzyme conjugate in 1% BSA/PBS), 25 ml per well - dilution as per manufacturer's instructions.

9. Wrap and incubate at 37oC for 1 hour.

10. Shake off and wash 10 X with PBS/Tween-20 (0.05%) or distilled water.

11. Add substrate solution per well.

12. Develop at room temp., reading at 15 min, 30 min and 1 hour.

Read More

Question
Answer


The lab let me know that they used a standard ELISA protocol with 50 microgram per well as a starting concentration with serial dilutions. The urea did not interfere.

Read More

Answer

Thank you for contacting us. In theory the protein ab49029 should react with antibidy ab124526, since the protein is full length Hepatitis C Virus NS4 protein. However, this has not been tested and there is a possibility the tag could prevent the antibody from binding. The ratio of biotin/IgG is 11/4 for the antibody.


I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More

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

Sign up