Overview

  • Product name
  • Description
    Goat polyclonal to Rotavirus
  • Host species
    Goat
  • Tested applications
    Suitable for: ICC/IF, Conjugationmore details
  • Species reactivity
    Reacts (>90%) with Human rotaviruses.
  • Immunogen

    Bovine Nebraska Calf Diarrhea Virus.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer
    pH: 7.20
    Preservative: 0.1% Sodium azide
    Constituent: 0.0268% PBS
  • Concentration information loading...
  • Purity
    Protein G purified
  • Purification notes
    >95% pure.
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab20036 in the following tested applications.

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

Application Abreviews Notes
ICC/IF Use at an assay dependent dilution.
Conjugation Use at an assay dependent dilution.

Target

  • Relevance
    Rotaviruses, members of the family Reoviridae, are a major cause of diarrhoea in young mammals. Rotavirus infections also result in economic losses in agriculture due to diarrhoea in calf, pig, sheep, and poultry rearing. Diarrhoea (or scours) due to the rotavirus Nebraska Calf Diarrhea Virus can affect calves up to 30 days of age or older. Diarrhoea begins 2 to 3 days after exposure. Diagnosis is by history, lesions (ulcers on the tongue, lips, and mouth) and diagnostic laboratory tests. Mortality rates may be as high as 50 percent, depending on the secondary bacteria present. Human rotaviruses, the major aetiological agents of severe infantile diarrhoea worldwide, display surprisingly diverse and complex serotypic specificities. Rotaviruses are 70 nm, non enveloped viruses comprised of a triple layered protein capsid; Outer capsid proteins are VP4 and VP7, Inner capsid -VP6 and Core -VP2. The immunity acquired from exposure to rotavirus appears to be type specific following initial infection; therefore, multiple serotypes of rotavirus mean multiple opportunities for infection. The combination of animal reservoirs for the virus and rotavirus gene reassortment provides the potential for dramatic genetic shifts (similar to influenza virus) which could give rise to altered host ranges and viral virulence.
  • Alternative names
    • Major inner capsid protein VP6 antibody
    • NSP4 antibody
    • VP6 antibody

References

This product has been referenced in:
  • Yang Y  et al. Immunogenicity and virus-like particle formation of rotavirus capsid proteins produced in transgenic plants. Sci China Life Sci 54:82-9 (2011). WB . Read more (PubMed: 21104033) »
See 1 Publication for this product

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Answer

I have been in contact with the lab regarding the issues which you have been experiencing with ab20036. Per the lab, this product does stain the cytoplasma of infected cells, but it is not a strong antibody in IFA. The lab recommends using it at a 1:100 to 1:200 dilution. They also recommend fixing the cells with cold acetone as the 10% formalin may destroy some of the epitopes.

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Answer

This is a polyclonal antibody that was raised using the whole virus, so it reacts with a number of the viral proteins. In ICC/IF, it gives cytoplasmic staining.

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Answer

Thank you for taking the time to contact us. I am sorry to hear you have some concerns regarding the results from this antibody.

I can confirm the immunogen used to producethis antibodyis Rotavirus NCDV (Nebraska Calf Diarrhea Virus). I'msorry wedo not have the amino acid sequence available for the complete virus.


I would like to reassure you that this antibody is tested and covered by our 6 month guarantee for immunofluorescence and conjugation. In the event that a product is not functioning in the tested applications and species cited on the product data sheet (and the problem has been reported within 6 months of purchase), we will be pleased to provide a credit note, free of charge replacement or refund.

I am sorry to confirm this antibody is regrettably not tested and covered by our guaranteed in western blot. Could you confirm if it has been tried in ICC-IF which is covered by our guarantee.

I would like to investigate this particular case further for you, and also obtain more information for our quality monitoring records. In order to proceed with this, I have enclosed a technical questionnaire below. I would appreciate if you could complete this. It will help you put the information we require together very easily. I have filled in the information already provided.

I would appreciate if you could also provide an image which would help us to assess the results.

After reviewing the details, there are somerecommendations I can suggestto consider:

1. I can suggest to use a lysis buffer such as RIPA to prepare the sample. This should provide a more suitable protein preparation.

2. Has the transfer to the membrane and quality of the sample been assessed with a loading control?

3, Has the amount of virus in the sample been assessed by any other methods?

Thank you for your time and cooperation. We look forward to receiving the completed questionnaire.



Order Details
Antibody code: ab20036

Problem
Choose: Non-specific band Multiple bands No signal or weak signal High background

Lot number

Purchase order number
or preferably Abcam order number:



General Information
Antibody storage conditions (temperature/reconstitution etc)


Description of the problem (high background, wrong band size, more bands, no band etc.)


Sample (Species/Cell extract/Nuclear extract/Purified protein/Recombinant protein etc.)


Sample preparation (Buffer/Protease inhibitors/Heating sample etc.)
PBS buffer with 0.02% Tween 20 for extraction including Sigma proteinase inhibitor cocktail.

Amount of protein loaded
loading 25-35 ug protein per lane

Electrophoresis/Gel conditions (Reducing or Non-reducing gel, % of the gel etc.)
12% SDS-PAGE then transfer to PVDF membrane


Transfer and blocking conditions (Buffer/time period, Blocking agent etc.)


Transfer 11 Volts, 2 hours

Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)
1:5000 dilution of ab20036 (4.5 mg/ml overnight incubation @ room temperature) followed by thrice 10 min washes with PBS-T

Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)
ab97120 (GR62374-2, donkey anti-Goat IgG HRP, 1:2500, 0.5 mg/ml, 2 hour incubation @ room temperature) and once more thrice 10 min washes with PBS-T before ECL detection

Detection method (ECL, ECLPlus etc.)
ECL

Positive and negative controls used (please specify)



Optimization attempts (problem solving)
How many times have you tried the Western?



Have you run a "No Primary" control?
Yes No

Do you obtain the same results every time?
Yes No
e.g. are the background bands always in the same place?


What steps have you altered?


Additional Notes:


Image:
We would appreciate if you are able to provide an image (including molecular weight markers) which would help us to asess the results.

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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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