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LiquiChip Kits for bead-based cytokine and kinase assays

The LiquiChip Human and Mouse 10-Cytokine Kits and Ser/Thr Kinase Kit provide fast and accurate quantification of a range of cytokines and serine/threonine kinases. LiquiChip 10-Cytokine Kits enable simultaneous multiplex assay of 10 different cytokines using bead-based xMAP technology. The LiquiChip Ser/Thr Kinase Kit enables highly sensitive assay of kinases without using radioactivity.

LiquiChip Kits offer:
  • Fast and sensitive assay on the LiquiChip workstation
  • Complete assay kits including beads, buffers, and detection reagents
  • Assay template software for fast startup
Fast and sensitive assay

Bead-based LiquiChip Cytokine and Kinase assays can be performed in a homogeneous mix-and-measure format with no wash steps, increasing throughput and reducing hands-on time. LiquiChip assay kits provide everything needed for the assay procedure including LiquiChip beads, buffers, and detection reagents enhancing sample-to-sample and plate-to-plate reproducibility. The multiplexing capability of the LiquiChip System enables comprehensive sample profiling in a single well, using a minimal amount of material.

Sensitive, multiplex human and mouse cytokine assays

LiquiChip Human and Mouse 10-Cytokine Kits enable fast, accurate, and simultaneous quantification down to picogram levels of 10 separate human or mouse cytokines in a single sample (Table 1). Antibodies that are highly specific for individual cytokines are immobilized on LiquiChip beads with different bead codes. Beads are added to samples, where cytokines bind to their respective antibody. Bead-bound cytokine s are detected using a mixture of biotinylated cytokine-specific monoclonal antibodies and StreptavidinPE (Figures 1 and 2). The specific bead code assigned to each of the 10 cytokines enables their unambiguous identification and quantification in the assay.

Table 1. Cytokines measured using LiquiChip 10-Cytokine Kits Sensitive and Highly Specific Detection of Cytokines Stimulation of Cytokines in Jurkat Cells Figure 1 Detection of cytokines using LiquiChip Cytokine Kits. Figure 2 Cultured Jurkat cells grown at 37C and 5% CO2 were exposed to 5 mM butyric acid for 18 hours. Cell lysates from treated and untreated cells were assayed using the LiquiChip Human 10-Cytokine Kit. The increase in fluorescent intensities due to treatment is shown. Kinase assay without radioactivity

The LiquiChip Ser/Thr Kinase Kit is based on the phosphorylation of myelin basic protein (MBP) immobilized on LiquiChip beads. After incubation with a sample, phosphorylation of serine and threonine residues in MBP is detected using antibodies specific for phosphoserine and phosphothreonine. These
antibodies are in turn detected using a biotinylated secondary antibody and streptavidinPE (Figure 3). The effect of kinase inhibitors can be measured by adding the inhibitor and a kinase to the assay. The LiquiChip Ser/Thr Kinase Kit enables quantification of numerous serine/threonine kinases, including JNK, p38-MAPK, SAPK 4, PKB-alph a, PKC, Erk-2, and GSK-. The LiquiChip assay procedure uses fluorescently labeled detection reagents to quantify kinase activity
with high sensitivity (Figure 4) and specificity, eliminating the need for radioactivity and its attendant handling problems.

Sensitive and Specific Kinase Detection Stimulation of Kinases in Jurkat Cells Figure 3 Detection of serine/threonine kinase activity using the LiquiChip Ser/Thr Kinase Kit. Figure 4 Cultured Jurkat cells grown at 37C and 5% CO2 were exposed to a range of kinase-inducing reagents for 18 hours. EGF, epidermal growth factor (concentration = 50 ng/ml in medium); PMA, phorbol 12-myristate 13-acetate (50 ng/ml); PHA, phytohemagglutinin (50 ng/ml); PMA+PHA (50 ng/ml each); BA, butyric acid (5 mM); LPS, lipopolysaccharide (250 ng/ml). Kinase activity was measured in singleplex LiquiChip Ser/Thr Kinase assays using undiluted lysates from treated and untreated cells. The increase in
fluorescent intensity due to treatment is shown.



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