BD™ Cytometric Bead Array (CBA) Human IgE Flex Set

The BD CBA Human IgE Flex Set must be used in conjunction with a BD CBA Human Soluble Protein Master Buffer Kit (Cat. No. 558264, 100 tests, or 558265, 500 tests), a flow cytometer, and the FCAP Array™ Software (Cat. No. 641488). Detailed instructions on the use of this product can be found in the manual for the BD CBA Human Soluble Protein Master Buffer Kit. The top standard point for the BD CBA Human IgE Flex Set is 500 ng/ml and should be prepared in the following way:

For single-plex Human IgE Flex Set experiments, make the top standard dilution by mixing 20µl of the Human IgE Standard with 980µl of Assay Diluent from the Human Soluble Protein Master Buffer Kit in a tube labeled Top Standard. Perform additional standard dilutions using the top standard dilution by following the instructions in the Human Soluble Protein Master Buffer Kit manual.

For multi-plex experiments including the Human IgE Flex Set, reconstitute the lyophilized standard proteins as directed in the Human Soluble Protein Master Buffer Kit manual, but use 20µl less Assay Diluent per ml used for reconstitution (e.g., if reconstituting lyophilized pellets in 1 ml of Assay Diluent, use 980µl instead). Transfer 980µl of the reconstituted proteins to a tube labeled Top Standard and add 20µl of the Human IgE Standard. Perform additional standard dilutions using the top standard dilution by following the instructions in the Human Soluble Protein Master Buffer Kit manual.

An example standard curve is shown in Figure 1.

The BD CBA Human IgE Flex Set should not be used in the same assay well with any non-BD CBA Human Soluble Protein Flex Set reagents (such as BD CBA Mouse Soluble Protein, Human Immunoglobulin or Cell Signaling Flex Sets). The BD CBA Human IgE Flex Set can not be used in the same assay well with any other Human immunoglobulin Flex Set assay or with the Human Ig Master Buffer Kit. For an updated assay compatibility chart for the BD CBA Human Soluble Protein Flex Sets, please refer to the BD CBA Flex Set System homepage at http://www.bdbiosciences.com/flexset.

Performance

Limit of Detection: The theoretical limit of detection is 0.07 ng/ml and was determined by evaluating the estimated result of the average MFI of the negative control (0 pg/ml, n=30) + 2 standard deviations.