Hispec Assay Diluent | N/A
HiSpec assay diluent used to dilute standards and samples in a sandwich ELISA for M. tuberculosis modified lipoprotein.
Image caption:
(A) Example schematic of a sandwich ELISA to detect TLP levels in 100μL infant serum samples from diagnosis of M. tuberculosis infection, using sandwich pair of Fab fragments 8 and 6. (B) Calibration curve of sandwich ELISA method showing linear correlation between antigen specific binding of a pairwise combination of TLP specific Fab fragments 6 and 8 (y-axis) to increasing concentration of TLP in a TLP spiked cord blood serum sample (x-axis). A lower detection limit of 4-5 ng/mL in serum is shown.
From: Yang X, Wipperman MF, Nachman S, Sampson NS.
Exploring the value of Mycobacterium tuberculosis modified lipoprotein as a potential biomarker for TB detection in children.
BMC Infect Dis. 2022 Feb 17;22(1):158.
doi: 10.1186/s12879-022-07140-9.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
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E ResetHispec Assay Diluent
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- Hispec Assay Diluent
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- Intended Use
- 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 Bio-Rad 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 - Product Form
- Ready to use assay diluent buffer
- Preservative Stabilisers
- 0.1% Proclin™ 300
- Regulatory
- For research purposes only
- Guarantee
- Guaranteed until date of expiry. Please see product label.
- Acknowledgements
- Proclin™ 300 is a trademark of The Dow Chemical Company (“Dow”) or an affiliated company of Dow.
This product is shipped at ambient temperature. It is recommended to aliquot and store at -20°C on receipt. When thawed, aliquot the sample as needed. Keep aliquots at 2-8°C for short term use (up to 4 weeks) and store the remaining aliquots at -20°C.
Avoid repeated freezing and thawing as this may denature the product. Storage in frost-free freezers is not recommended.
Avoid repeated freezing and thawing as this may denature the product. Storage in frost-free freezers is not recommended.
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 visit the antibody protocols page.
| Application Name | Verified | Min Dilution | Max Dilution |
|---|---|---|---|
| ELISA |  |
||
| 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.
- 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. - 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. - 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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References for Hispec Assay Diluent
-
Eriksson, O. et al. (2018) Pancreatic imaging using an antibody fragment targeting the zinc transporter type 8: a direct comparison with radio-iodinated Exendin-4.
Acta Diabetol. 55 (1): 49-57. -
Yang, X. et al. (2022) Exploring the value of Mycobacterium tuberculosis. modified lipoprotein as a potential biomarker for TB detection in children.
BMC Infect Dis. 22 (1): 158.
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