BD, 554611, BD Pharmingen™ Recombinant Human IL-10

Preparation And Storage: Preparation And Storage Store product at -80°C prior to use or for long term storage of stock solutions. Rapidly thaw and quick-spin product prior to use. Avoid multiple freeze-thaws of product. This preparation contains no preservatives, thus it should be handled under aseptic conditions.
Recommended Assay Procedures: Recommended Assay Procedures Upon initial thawing the product should be aliquoted into polypropylene microtubes and frozen at -80°C for future use.  Alternatively, the product can be diluted in sterile neutral buffer containing not less than 0.5 - 10 mg/mL carrier protein, such as human or bovine albumin, aliquoted and stored at -80°C.    For in vitro biological assay use, carrier-protein concentrations of ≥ 1 mg/mL are recommended.  For use as an ELISA standard carrier-protein concentrations of 5 - 10 mg/mL are recommended.  Failure to add carrier protein or store at indicated temperatures may result in a loss of activity.  The product should not be diluted to less than 50 μg/mL for long term storage.  Carrier proteins should be pre-screened for possible effects in each investigator's experimental system.  Carrier proteins may have an undesired influence on experimental results due to toxicity, high endotoxin levels or possible blocking activity. ELISA Standard :  Recombinant human IL-10 (Cat. No. 554611)  is useful as a quantitative standard for measuring human IL-10 protein levels in an IL-10 specific sandwich ELISA with Purified Rat Anti-Human IL-10 (Cat. No. 554705) as a capture antibody and Biotin Anti-Human and Viral IL-10 (Cat. No. 554499) as the detection antibody.  To obtain linear standard curves, investigators may want to consider using doubling dilutions of recombinant human IL-10 standard from 2,000 - 15 pg/mL to be included in each ELISA plate.  For measuring human IL-10 in serum or plasma, investigators are highly encouraged to use the BD OptEIA™ Human IL-10 ELISA Set (Cat. No. 555157) and the BD OptEIA™ Human IL-10 ELISA Kit (Cat. No. 550613). Bioassay : Investigators are advised that the Bioassay application is not routinely tested for this material and are highly encouraged to both titrate this material and include appropriate controls in relevant experiments.  An  activity range of 0.5 - 3.3 x 10^7 units/mg, encompassing an ED50= 0.3 - 2 ng/mL, has previously been reported using MC/9 as indicator cells for proliferation, with a unit defined as the amount of material needed to stimulate a half-maximal response at cytokine saturation. Blocking :  Recombinant Human IL-10 (Cat. No. 554611) can be useful as a blocking control for flow cytometric analysis when used with fluorochrome conjugated antibodies, such as PE-conjugated JES3-9D7 (Cat. No. 554498).  Investigators are advised that the blocking application is not routinely tested for this material.  Intracellular cytokine staining techniques and use of blocking controls are described in detail by C.Prussin and D.Metcalfe.