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
- Fast and sensitive assay on the LiquiChip workstation
- Complete assay kits including beads, buffers, and detection reagents
- Assay template software for fast startup
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.
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
Sensitive and Highly Specific Detection of Cytokines
Stimulation of Cytokines in Jurkat Cells
Figure 1 Detection of cytokines using LiquiChip Cytokine
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.
Source:Page: All 1 2 3 Related biology technology :1
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