BPHRTK01  Widescreen^® BeadPlex^® Human Receptor Tyrosine Kinase Panels

BeadPlex^® Human Receptor Tyrosine Kinase (RTK) Total Panel and
BeadPlex^® Human Receptor Tyrosine Kinase (RTK) Phosphotyrosine (pTyr) Panel

xMAP^® technologies is a multiplex bead-based assay platform that allows for sensitive, precise quantification of multiple analytes within a single sample. For the detection of proteins or peptides, the assay kits often utilize the familiar sandwich or competitive immunoassay formats (e.g. ELISA). Unlike ELISA though, the target analytes are captured on differentially fluorescently-dyed beads. This differential allows for the detection of multiple analytes from the same sample in the same well. The BeadPlex^® assays are run by combining the sample(s) with the capture antibody-coated beads. The specific analytes from the sample are captured on the beads and are then subsequently identified with detection antibodies and a fluorescent label using an appropriate instrument. A major advantage over traditional protein quantification methods (such as ELISA) is the capacity for multiplexing, as bead-based assays allow simultaneous quantification of multiple analytes in a small sample volume.

Panel Overview


Receptor tyrosine kinases (RTKs) are critical regulators of numerous cell signaling pathways and have been implicated in various disease states. Ligand binding to the extracellular domain of transmembrane RTKs triggers receptor dimerization and autophosphorylation of an intracellular kinase domain. This event ultimately triggers activation of downstream pathway proteins via phosphotyrosine-SH2 domain interactions. The BeadPlex Human Receptor Tyrosine Kinase (RTK) Total Panel allows for simultaneous quantification of up to ten (10) human receptor tyrosine kinases: EGFR, HER-2, HER-3, HER-4, HGFR, IGF-1R, InsR, PDGFR-β, Tie-2, and VEGFR2, regardless of their phosphorylation state. The BeadPlex Human Receptor Tyrosine Kinase (RTK) Phosphotyrosine (pTyr) Panel allows for the qualitative detection of those same receptors in either their inactive or active state. Both panels can also be used in companion, but not multiplexed, with each other for normalizing the active RTK content of samples. By normalizing the signal from the pTyr assay to the RTK quantity determined by the RTK total assay, end-users can accurately determine the activation states of those aforementioned receptors.

The BeadPlex Human Receptor Tyrosine Kinase (RTK) Total and Phosphotyrosine (pTyr) Panel workflow is shown in the diagram at the bottom of this page. If expressed in the cell line of interest, individual RTK proteins may be present in either their inactive or active states depending on availability of the RTK ligand. Cellular protein samples are prepared by gentle membrane extraction. Extracts can be split and analyzed for the total RTK contents with the BeadPlex Human Receptor Tyrosine Kinase (RTK) Total Panel (EMD Chemicals, Inc., Cat. No. BPHRTK01-10TOT) or the active RTK contents with the BeadPlex Human receptor Tyrosine Kinase (RTK) phosphotyrosine (pTyr) Panel (EMD Chemicals, Inc., Cat. No. BPHRTK01-10PY).

These novel multi-analyte profiling kits provide a deeper view of the following Receptor Tyrosine Kinases:

• Epidermal Growth Factor Receptor (EGFR)
• Human Epidermal Growth Factor Receptor 2 (HER-2)
• Human Epidermal Growth Factor Receptor 3 (HER-3)
• Hepatocyte Growth Factor Receptor (Met)
• Insulin-like Growth Factor 1 Receptor (IGF-1R)
• Insulin Receptor (InsR)
• Platelet-Derived Growth Factor Receptor-beta (PDGFR-β)
• Tyrosine Kinase with Immunoglobulin and EGF Repeats 2 (Tie-2)
• Vascular Endothelial Growth Factor Receptor 2 (VEGFR2)

Panel Components

Storage: All components of the BeadPlex Human Receptor Tyrosine Kinase (RTK) Total Panel and the BeadPlex Human Receptor Tyrosine Kinase (RTK) Phosphotyrosine (pTyr) Panel are shipped at 2 to 8°C. Upon receipt, store all kit components at 2 to 8°C. Do Not Freeze.

The BeadPlex Human Receptor Tyrosine Kinase (RTK) Total Panel is available in two kit configurations:

• Pre-mixed Complete Kit
• Mix and Match Format

For utmost convenience, end-users may order the BeadPlex Human Receptor Tyrosine (RTK) Total Panel Complete Kit (EMD Chemicals, Inc., Cat. No. BPHRTK01-10TOT). This kit contains all respective capture beads and all the components in the BeadPlex Human Receptor Tyrosine (RTK) Total Base Kit. Desired capture beads are already pre mixed, but must be diluted accordingly before running an assay. A Complete Kit contains enough components to assay one 96-well plate.

For maximum flexibility, end-users may configure their own customized multiplex assay of up to ten specified analytes through the Mix and Match Format. This format consists of a BeadPlex Human Receptor Tyrosine Kinase (RTK) Total Panel Base Kit (EMD Chemicals, Inc., Cat. No. BPHRTK01-TBASE) and a customized choice of any or all analyte-specific capture beads as listed in Technical Bulletin (TB548). In the Mix and Match Format, desired capture beads will need to be mixed and diluted accordingly prior to running an assay. The Base Kit and selected components contain enough reagents and buffers to assay one 96-well plate.

Similarly, the BeadPlex Human Receptor Tyrosine Kinase (RTK) Phosphotyrosine (pTyr) Panel is also available in the two kit configurations:

• Pre-mixed Complete Kit
• Mix and Match Format

For utmost convenience, end-users may order the BeadPlex Human Receptor Tyrosine (RTK) Phosphotyrosine (pTyr) Panel Complete Kit (EMD Chemicals, Inc., Cat. No. BPHRTK01-10PY). This kit contains all respective capture beads and all the components in the BeadPlex Human Receptor Tyrosine (RTK) Phosphotyrosine (pTyr) Base Kit. Desired capture beads are already pre mixed, but must be diluted accordingly before running an assay. A Complete Kit contains enough components to assay one 96-well plate.

For maximum flexibility, end-users may configure their own customized multiplex assay of up to ten specified analytes through the Mix and Match Format. This format consists of a BeadPlex Human Receptor Tyrosine Kinase (RTK) Phosphotyrosine (pTyr) Panel Base Kit (EMD Chemicals, Inc., Cat. No. BPHRTK01-YBASE) and a customized choice of any or all analyte-specific capture beads as listed in Technical Bulletin (TB549). In the Mix and Match Format, desired capture beads will need to be mixed and diluted accordingly prior to running an assay. The Base Kit and selected components contain enough reagents and buffers to assay one 96-well plate.