Invitrogen™ Antibody Isotyping 6-Plex Rat ProcartaPlex™ Panel

Invitrogen™ Antibody Isotyping 6-Plex Rat ProcartaPlex™ Panel

The Rat Antibody Isotyping 6-Plex ProcartaPlex Panel enables determination of antibody heavy chain subclasses using Luminex xMAP technology. Antibody isotyping is a critical and beneficial tool in hybridoma development. Simultaneous detection of these targets can help explore analysis of immunoglobulin isotypes during and after hybridoma development and to assess immune responses. The panel is provided in a ready-to-use format with individual vials of capture and detection reagents formulated at 1X concentration, requiring less pipetting and experimental setup. It is not combinable with simplexes or other panels.

ProcartaPlex preconfigured panels are extensively tested for analyte combinability, interference, and cross-reactivity to provide the highest level of validation and precision. All ProcartaPlex panels are supplied with the necessary reagents to perform the assay.

ProcartaPlex assays utilize Luminex xMAP (multianalyte profiling) technology for the simultaneous detection and quantitation of up to 65 protein targets in a single 25–50 μL sample—from plasma, serum, cell culture supernatants, and other bodily fluids.

The Luminex beads in the ProcartaPlex assay are internally dyed with precise proportions of red and infrared fluorophores to create spectrally unique signatures that can be identified by the Luminex xMAP detection systems (e.g., Luminex 200, FLEXMAP 3D, and MAGPIX). Similar to a sandwich ELISA, the ProcartaPlex assay uses matched antibody pairs to identify the protein of interest. In a multiplexed assay, each spectrally unique bead is labeled with antibodies specific for a single target protein, and bound proteins are identified with biotinylated antibodies and streptavidin–R-phycoerythrin (RPE). The conjugation of protein-specific antibodies to a distinct bead allows for analysis of multiple targets in a single well.
The most significant difference between a ProcartaPlex assay and ELISA is that the capture antibody in the ProcartaPlex assay is conjugated to a bead and not adsorbed to the microplate well, so the ProcartaPlex assay reagents are free-floating in the solution. For detection, the Luminex 200 instrument, for example, contains two lasers, one to distinguish the spectral signature of each bead and the second to quantify the amount of RPE fluorescence, which is proportional to the amount of protein present in the sample. ProcartaPlex multiplex assays can profile more target proteins using significantly less sample in the same time that it takes to perform a traditional sandwich ELISA.

Target list [bead region]:

• IgG1 [25], IgG2a [28], IgG2b [34], IgG2c [44], IgA [61], IgM [48]

Recommended sample pre-dilution: Serum, Plasma: 1:40000; Cell culture supernatant: 1:200

Features

• Reproducible, reliable results—validated as a panel to the highest industry standard, including protein target combinability and cross-reactivity testing.
• More results per sample—measure multiple protein targets simultaneously in a single 25–50 µL sample.
• Well-established Luminex technology highly referenced multiplexing platform for protein detection and quantitation.

Contents

• 2 vials Rat Isotyping Standard Mix (lyophilized)
• 1 vial Capture Bead Mix (1X)
• 1 vial Biotinylated Detection Antibody Mix (50X)
• 1 bottle Reading Buffer (1X)
• 1 bottle Wash Buffer (10x)
• 1 bottle Universal Assay Buffer (10x)
• 1 bottle Detection Antibody Diluent (1X)
• 8-tube strip
• Adhesive film
• Flat-bottom 96-well plate, black