Human Chemokine Panel
Size: 100 Tests
Assay Range: 3.9-1,000 Units/mL
Beads-based assays provide a method of capturing a soluble analyte or set of analytes with beads of known size and fluorescence, making it possible to detect analytes using flow cytometry. Each capture bead in the kit has been conjugated with a specific antibody. The detection reagent provided in the kit is a mixture of phycoerythrin (PE)–conjugated antibodies, which provides a fluorescent signal in proportion to the amount of bound analyte. When the capture beads and detection reagent are incubated with an unknown sample containing recognized analytes, sandwich complexes (capture bead + analyte + detection reagent) are formed. These complexes can be measured using flow cytometry to identify particles with fluorescence characteristics of both the bead and the detector.
Five bead populations with distinct fluorescence intensities have been coated with capture antibodies specific for CXCL8/IL-8, CCL5/RANTES, CXCL9/MIG, CCL2/MCP-1, and CXCL10/IP-10. The five bead populations are mixed together to form the bead array that is resolved in a red channel (eg, FL3 or FL4) of a flow cytometer. During the assay procedure, you will mix the chemokine capture beads with the recombinant standards or unknown samples and incubate them with the PE conjugated detection antibodies to form sandwich complexes. The intensity of PE fluorescence of each sandwich complex reveals the concentration of that cytokine. After acquiring samples on a flow cytometer, use FCAP Array™ software to generate results in graphical and tabular format.
Contents This kit contains the following components sufficient for 80 tests.
Human Chemokine Capture Beads (A1–A5): An 80-test vial of each specific capture bead (A1–A5). The specific capture beads, having discrete fluorescence intensity characteristics, are distributed from brightest (A1) to dimmest (A5).
Cytometer Setup Beads (D): A 30-test vial of setup beads for setting the initial instrument PMT voltages and compensation settings is sufficient for 10 instrument setup procedures. The Cytometer Setup Beads are formulated for use at 50 µL per test.
Antibody and standard reagents
Human Chemokine PE Detection Reagent (B): An 80-test vial of PE-conjugated anti-human CXCL8/IL-8, CCL5/RANTES, CXCL9/MIG, CCL2/MCP-1, and CXCL10/IP-10 antibodies, formulated for use at 50 µL
Human Chemokine Standards: Two vials containing lyophilized recombinant human proteins. Each vial should be reconstituted in 4.0 mL of Assay Diluent to prepare the top standard.
PE Positive Control Detection: A 10-test vial of PEconjugated antibody control formulated for use at 50 µL per test. This reagent is used with the Cytometer Setup Beads to set the initial instrument compensation settings.
FITC Positive Control Detection: A 10-test vial of FITC-conjugated antibody control formulated for use at 50 µL per test. This reagent is used with the Cytometer Setup Beads to set the initial instrument compensation settings.
Wash Buffer (F): A 130-mL bottle of phosphate buffered saline (PBS) solution (1X), containing protein and detergent, used for wash steps and to resuspend the washed beads for analysis. Assay Diluent (G): A 30-mL bottle of a buffered protein solution (1X) used to reconstitute and dilute the Human Chemokine Standards and to dilute the unknown samples. Note: Source of all serum proteins is from USDAinspected abattoirs located in the United States.
Store all kit components at 2 to 8°C. Do not freeze.
Components contain sodium azide. Sodium azide yields a highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discharging to avoid accumulation of potentially explosive deposits in plumbing.
Human Chemokine Standards contains 0.02% (w/w) of a CMIT/MIT mixture (3:1), which is a mixture of: 5-chloro-2-methyl-4-isothiazolin-3-one [EC No 247-500-7] and 2-methyl-4-isothiazolin-3-one [EC No 220-239-6] (3:1).
• May cause an allergic skin reaction.
• Wear protective gloves / eye protection.
• Wear protective clothing.
• Avoid breathing mist/vapours/spray.
• If skin irritation or rash occurs: Get medical advice/attention.
• IF ON SKIN: Wash with plenty of water.
• Dispose of contents/container in accordance with local/regional/national/international regulations.
This product is for research only, not for diagnosis purpose.