Bead-Based Immunoassays | BD Biosciences

Overview

The BD^® Cytometric Bead Array (CBA) Solutions measure a variety of soluble and intracellular proteins, including cytokines, chemokines, growth factors and phosphorylated signaling proteins using flow cytometry.

The solutions:

Enable analysis of up to 30 proteins using just 25 to 50 μL of sample
Are compatible with almost any standard flow cytometer without the need to purchase expensive single-purpose equipment
Are available in preconfigured kits or in an open configurable format for unique needs
- BD^® CBA Kits—preconfigured for simple set up and to minimize the need for optimization of routine panel
- BD^® CBA Flex Sets—open and configurable method of detection to build your own multiplexes for protein quantitation as low as 10 pg/mL
- BD^® CBA Enhanced Sensitivity Flex Sets—capable of detecting cytokine concentrations as low as 0.274 pg/mL
- BD^® CBA Functional Beads—allow you to conjugate your own antibody for your protein of interest

FEATURES

BD^® CBA Kits are preconfigured by functional areas of biology (for example, Th1/Th2 or inflammatory cytokines) and allow simultaneous measurement of up to seven analytes

The kits provide consistent results with routine panels

The kits:

Provide preconfigured panels for ease of use and enable multiplex analysis of complex biological samples on a flow cytometer
Use capture beads that contain unique amounts of a single red dye, whose unique spectral properties enable analysis of samples on single 488-nm laser or dual-laser (488-nm or 532-nm and 633-nm) flow cytometers
Are ready to use and complete with all the buffers and reagents necessary to analyze 80 samples
Offer assays that are stringently developed for ease of use and rapid data analysis and that use antibody pairs chosen for their sensitivity, consistency and quality
Contain antibody pairs that are evaluated for dynamic range, sensitivity and parallel titration curves to native biological samples
Provide assay diluent and wash buffers that have been formulated to reduce detrimental effects of serum and plasma proteins on assay performance
Use FCAP Array™ Analysis Software, which gates on each individual bead population and determines the median fluorescence intensity (MFI) for each analyte in the array and generates a standard curve

Refer to the brochure for FCAP Array™ Software parameters.

BD^® CBA Flex Sets allow simultaneous measurement of up to 30 analytes on a flow cytometer equipped with either a 488-nm or 532-nm laser, and a 633-nm laser

The flex sets are open and configurable to maximize flexibility and to make it easy to create multiplex assays

The sets:

Provide specificities for the detection of human, mouse or rat cytokines, chemokines and growth factors; human immunoglobulins; and cell signaling assays for the detection of phosphorylated cell signaling proteins
Offer assays formulated for mixing into panels of up to 30-plex and are all sold individually to provide a highly flexible system. All products are available off the shelf so custom orders are not required.
Contain a unique Master Buffer Kit for each product area (Human Soluble Protein, Mouse/Rat Soluble Protein, and Human Immunoglobulin). All assays within each product area have been verified for performance in single-plex and in multiplexed scenarios to ensure consistency between assay types.
Make it easy to build a multiplex by following five simple steps. The finished assay can be acquired on a variety of dual-laser flow cytometers and analyzed using FCAP Array™ Software or directly with FlowJo™ v10 Software using a downloadable plug-in.

Refer to the brochure for FCAP Array™ Software parameters.

BD^® CBA Enhanced Sensitivity Flex Sets allow you to detect <1 pg/mL of cytokines and get quantitative results in samples that were below the level of detection using standard BD^® CBA Flex Sets

The kits offer a reliable, flexible method for the quantitative detection of multiple proteins in a single serum, plasma or tissue culture supernatant sample

The kits:

Provide the ability to detect protein as low as 0.274 pg/mL, more than a 35-fold improvement compared to most bead assays that can detect down to 5–10 pg/mL
Include assays for 10 different cytokines involved in inflammation and leucocyte differentiation that can be combined to create a multiplexed panel
Provide more data in less time from a single sample by allowing you to analyze up to 10 proteins with just 25 to 50 μL of sample. Other methods such as ELISAs require a similar amount of sample to analyze only one protein.
Are formulated for combination in multiple configurations and sold individually to provide a highly flexible system. A dedicated Master Buffer Kit contains all the buffers needed for the assay.
Offer assays that have been characterized for performance in single-plex and multiplex scenarios to ensure consistency across assay types. The antibody pair in each assay is evaluated for dynamic range, sensitivity and parallel titration to native biological samples.
Provide optimized assay protocols to minimize background and maximize signal while using a two-step detection protocol
Use FCAP Array™ Analysis Software, which gates on each individual bead population and determines the median fluorescence intensity (MFI) for each analyte in the array and generates a standard curve

Refer to the brochure for FCAP Array™ Software parameters.

BD^® CBA Functional Beads allow you to conjugate your own antibody for your protein of interest

The beads provide additional options to enhance flexibility

The unconjugated beads:

Allow conjugation of antibody or protein in less than four hours using common laboratory supplies along with the buffers in the BD^® CBA Functional Bead Conjugation Buffer Kit
Provide an ideal platform for converting existing ELISAs into bead-based immunoassays that can be mixed with our portfolio of BD^® CBA Flex Set assays
Enable the creation of novel multiplex panels with multiple bead positions
Offer the ability to prepare up to 1,000 tests in a single reaction, ensuring consistency across a large number of tests