Pro5® MHC Class I Pentamers and Related Products

Flexible Labeling Options

Unlabeled Pentamer

Pro5 Fluorotag

For optimum flexibility Pentamers may be ordered labeled with R-PE, APC or biotin, or unlabeled with an additional Pro5® Fluorotag or Pro5® Biotag. Directly labeled Pentamers are stored at 4°C and are quick and convenient to use. They are especially valuable in the analysis of a single donor sample for multiple peptide specificities. A single donor sample may be simultaneously stained and analyzed for multiple specificities by double or triple staining with labeled Pentamers. This generates a more complete profile of the immune response whilst reducing the total number of cells required for staining. Any combination of R-PE-labeled, APC-labeled and biotin-labeled Pentamers may be used for this purpose.

Unlabeled Pentamers may be stored at -80°C and are guaranteed for 12 months under such conditions, making them perfect for long-term studies. Internal validation of a large batch of Pentamer at the beginning of a study saves time by not having to re-qualify new reagent batches every few months. Unlabeled Pentamers offer the user added staining flexibility, in that the same complex may be used with a number of different fluorochromes. This feature makes experimental design more adaptable, especially when choosing antibodies to cell surface markers or intracellular proteins that are only available in certain colors.

Pro5® Pentamers are available in 50, 150 and 500 test quantities. One test of Pentamer is sufficient to stain approximately 1-2 x 106 PBMCs. Pentamers can be supplied unlabeled or labeled, with a choice of R-PE, APC or biotin. R-PE or APC fluorescent Pro5® Fluorotags and Pro5® Biotag are available separately for use with unlabeled Pentamers in corresponding quantities of 50, 150 and 500 tests.

Staining Protocols for Labeled and Unlabeled Pro5® MHC Class I Pentamers

Comprehensive staining protocols for Pro5® Pentamers are available in the Pro5® MHC Pentamer Handbook and on the Protocols page

Download Pentamer Handbook

 

Flow diagram of labeled and unlabeled Pentamer staining protocols

 

Pro5® MHC Class I Pentamer and Anti-CD8 Kits

Pro5® Pentamer analysis should be carried out with co-staining of the CD8+ T cell population. All Pentamers are optionally available in a kit with a FITC-labeled, anti CD8 monoclonal antibody (human or mouse). The antibody clone supplied in the kit has been tested for superior performance in conjunction with Pentamer products, and is provided as a separate aliquot. This enables the user to titrate the antibody in order to find the optimum working dilution.  There are over 250 different Pentamer specificities available in our catalog, encompassing epitopes for cancer and infectious diseases such as CMV, EBV, HBV, HCV, HPV, Influenza and many more. List of catalogue Pentamers.

Custom Pro5® MHC Class I Pentamers

If your chosen specificity is not in our range, custom Pro5® Pentamers are available in the same format as catalog items. Our technical expertise and experience making thousands of different MHC-peptide complexes means that we are able to provide custom reagents efficiently and with a high rate of success.  Custom Pentamers are usually subject to a non-refundable set-up charge, which will be applied once for each item. The set-up charge will not be applied to repeat orders of the same Pentamer. The price for a custom Pentamer includes the synthesis of the specific custom peptide. ProImmune will only proceed with the synthesis of a custom Pentamer if we are confident that the peptide will bind sufficiently to the relevant MHC allele. Usually synthesis will proceed if the affinity score is 21 or higher on www.syfpeithi.com. Lower scoring peptides will be considered if sufficient supporting evidence is available for binding.

Matched Peptides

ProImmune provides matched peptides for all catalog peptide specificities and custom Pro5® Pentamers. Matched peptides are provided lyophilized in 1 or 2 mg quantity, and at a guaranteed purity of >70% (immunograde). They are suitable for in vitro T cell stimulation experiments and can be purchased at a much-reduced cost compared to a separate standard custom peptide synthesis. Matched peptides are dispatched in less than one week.

Links to further information about Pro5® MHC Class I Pentamers

Introduction to Pentamers

Pentamer Applications

Biotin-labeled Pentamers

List of Catalog Pro5® Pentamers

Custom Pro5® Pentamers

Technical Webcasts about Pro5® Pentamers

Publications

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