HISPEC assay diluent works to reduce cross reactivity, non-specific binding and matrix effects in immunoassays such as ELISA, EIA, Western blotting, immuno-PCR, protein arrays, multianalyte immunoassays and immunohistochemistry.
The effect of HISPEC assay diluent is dependent on assay system and the antibodies used. HISPEC assay diluent is used instead of a sample buffer or antibody dilution buffer within the immunoassay protocol.
HISPEC assay diluent is unsuitable for blocking of surfaces. For blocking of surfaces we recommend one of the blocking solutions within the AbD Serotec range – see “Useful reagents” below.
Examples of use:
ELISA: use as a dilution buffer for the specimen and for the detection antibodies
Western blotting: use as dilution buffer for primary and secondary antibodies
Immunohistochemistry: use as dilution buffer for primary and secondary antibodies
Protein arrays: use as dilution buffer for specimen and for the detection antibodies
Application
This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visitwww.abdserotec.com/protocols.
Application Name
Yes
No
Not Determined
Suggested Dilution
Immunoassay
Where this product has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own system using appropriate negative/positive controls.
Storage
Store at +4oC or at -20oC if preferred.
Storage in frost-free freezers is not recommended.
This product should be stored undiluted. Avoid repeated freezing and thawing.
Shelf Life
12 months from date of despatch at -20oC or 6 months at 2-8oC.
Instructions For Use
The buffer should be mixed thoroughly immediately before use.
Dilution of the samples:
The standards and samples for ELISA and protein arrays can be diluted with HISPEC assay diluent at 1:2 or higher. Standards and samples should be treated identically.
Dilution of antibodies:
Antibodies can be diluted with HISPEC assay diluent in a user-defined manner, depending on the recommendation of the antibody supplier.
Expected results:
In some cases a reduction in assay signal intensity may be observed when using HISPEC assay diluent. This does not result in a reduction in assay sensitivity due to the accompanying reduction in non-specific binding.
Signal intensity may be increased in some assay systems by use of higher concentrations of antibodies, or by dilution of the HISPEC assay diluent in distilled water (1:2) prior to use. However, these adjustments may also result in some associated increase in non-specific binding.
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