Product Information
Product Name: SpeedyPLEX Human/Non-Human Primate Th1/Th2 Cytokine (6-plex) CBA Assay Panel
Sample type: Serum, Plasma and Cell culture supernatant
Test number: 96 tests
Target name | Standard Curve Range (pg/mL) | Sensitivity (pg/mL) |
| IL-2 | 1.37-1000 | 0.03 |
| IL-4 | 2.74-2000 | 0.08 |
| IL-5 | 2.74-2000 | 0.36 |
| IL-6 | 2.74-2000 | 0.10 |
| TNF-alpha | 2.74-2000 | 0.35 |
| IFN-gamma | 2.74-2000 | 0.02 |
Product Components
Name | Size | Quantity | Storage |
Microplate - 96-Well Plate (Flat bottom)1 | 8 holes × 12 strips | 1 plate | Unopened Kit: Store all reagents at 2-8°C for 12 months. Please use a new standard for each assay. |
Microplate - 96-Well Plate (V-bottom)2 | 8 holes × 12 strips | 1 plate | |
Setup Beads Cocktail* | 500 uL/vial | 1 vial | |
PE Control Beads* | 500 uL/vial | 1 vial | |
Protein Standard Cocktail; lyophilized | - | 2 bottles | |
Capture Beads Cocktail (50x)* | 120 uL/vial | 1 vial | |
Biotinylated Detection Antibody Cocktail (50x)* | 120 uL/vial | 1 vial | |
Sample Diluent PT 3-ef | 30 mL/bottle | 1 bottle | |
Detection Diluent | 30 mL/bottle | 1 bottle | |
Additional Diluent ATK00005 | 6 mL/bottle; Only for serum and plasma samples | 1 bottle | |
Wash Buffer Concentrate (20x) | 30 mL/bottle | 1 bottle | |
Streptavidin-PE (50x)* | 120 uL/vial | 1 vial | |
Plate Cover Seals | - | 4 pieces |
1: Microplate - 96-Well Plate (Flat bottom) match with Hand-held microplate magnet or platewasher with a magnetic platform in the wash step.
2: Microplate - 96-Well Plate (V-bottom) matches with centrifugal machine in the wash step.
* Centrifugation immediately before use.
Sample values
Human serum - Twelve samples were evaluated for detectable levels of IL-2, IL-4, IL-5, IL-6, TNF-alpha and IFN-gamma in this assay, the concentrations measured are shown below.
Analyte | Range (pg/mL) | No. of Detectable | Mean (pg/mL) | % of Detectable |
IL-2 | ND | 0 | ND | 0% |
IL-4 | ND | 0 | ND | 0% |
IL-5 | ND-11.41 | 8 | 2.18 | 67% |
IL-6 | 1.83-20.55 | 12 | 10.43 | 100% |
TNF-alpha | 1.66-5.93 | 12 | 3.73 | 100% |
IFN-gamma | 0.02-3.49 | 12 | 0.89 | 100% |
ND*=Non-detectable
Cynomolgus monkey serum - Eight samples were evaluated for detectable levels of IL-2, IL-4, IL-5, IL-6, TNF-alpha and IFN-gamma in this assay, the concentrations measured are shown below.
Analyte | Range (pg/mL) | No. of Detectable | Mean (pg/mL) | % of Detectable |
IL-2 | 0.03-16.24 | 8 | 2.1 | 100% |
IL-4 | ND | 0 | ND | 0% |
IL-5 | ND-5.63 | 2 | 0.82 | 25% |
IL-6 | ND | 0 | ND | 0% |
TNF-alpha | ND-2.62 | 6 | 0.76 | 75% |
IFN-gamma | 3.34-15.45 | 10 | 9.71 | 100% |
ND*=Non-detectable
Cell culture supernatant - Human peripheral blood mononuclear cells (PBMC) (1 x 106 cells/mL) were cultured in DMEM supplemented with 10% fetal bovine serum, 5 μM β-mercaptoethanol, 2 mM L-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin sulfate. Cells were cultured unstimulated or stimulated with 10 μg/mL PHA for 1 day. Aliquots of the cell culture supernatants were removed and assayed for levels of IL-2, IL-4, IL-5, IL-6, TNF-alpha and IFN-gamma.
IL-2 (pg/mL) | IL-4 (pg/mL) | IL-5 (pg/mL) | IL-6 (pg/mL) | TNF-alpha (pg/mL) | IFN-gamma (pg/mL) | |
Unstimulated (1 day) | 11.97 | ND | 4.58 | 6.84 | 13.10 | 9.21 |
Stimulated (1 day) | 12,140.19 | 274.62 | 323.14 | 14,148.25 | 7,270.43 | 29,671.91 |
ND*=Non-detectable
Cynomolgus monkey peripheral blood mononuclear cells (PBMC) (1 x 106 cells/mL) were cultured in DMEM supplemented with 10% fetal bovine serum, 5 μM β-mercaptoethanol, 2 mM L-glutami-ne, 100 U/mL penicillin, and 100 μg/mL streptomycin sulfate. Cells were cultured unstimulated or stimulated with 10 μg/mL PHA for 3 days. Aliquots of the cell cu-lture supernatants were remov-ed and assayed for levels of IL-2, IL-4, IL-5, IL-6, TNF-alpha and IFN-gamma.
IL-2 (pg/mL) | IL-4 (pg/mL) | IL-5 (pg/mL) | IL-6 (pg/mL) | TNF-alpha (pg/mL) | IFN-gamma (pg/mL) | |
Unstimulated (3 days) | ND | ND | ND | 534.5 | 102.9 | ND |
Stimulated (3 days) | 14.5 | 39.1 | 83.8 | 14,440.1 | 3,102.3 | 10,306.7 |
ND*=Non-detectable
Cynomolgus monkey peripheral blood mononuclear cells (PBMC) (1 x 106 cells/mL) were cultured in DMEM supplemented with 10% fetal bovine serum, 5 μM β-mercaptoethanol, 2 mM L-glutami-ne, 100 U/mL penicillin, and 100 μg/mL streptomycin sulfate. Cells were cultured unstimulated or stimulated with 50 ng/mL PMA and 1 μg/mL calcium Ionomycin for 1 day. Aliquots of the cell cu-lture supernatants were removed and assayed for levels of IL-2, IL-4, IL-5, IL-6, TNF-alpha and IFN-gamma.
IL-2 (pg/mL) | IL-4 (pg/mL) | IL-5 (pg/mL) | IL-6 (pg/mL) | TNF-alpha (pg/mL) | IFN-gamma (pg/mL) | |
Unstimulated (1 day) | ND | ND | ND | 5394.9 | 1307.8 | 27.5 |
Stimulated (1 day) | 55,397.1 | 1,340.7 | 764.0 | 7,528.6 | 9,897.4 | 289,641.9 |
ND*=Non-detectable
Cynomolgus monkey peripheral blood mononuclear cells (PBMC) (1 x 106 cells/mL) were cultured in DMEM supplemented with 10% fetal bovine serum, 5 μM β-mercaptoethanol, 2 mM L-glutami-ne, 100 U/mL penicillin, and 100 μg/mL streptomycin sulfate. Cells were cultured unstimulated or stimulated with PHA and IL-2 for 8 days, and PHA, PMA and Ca2+ for 1 day. Aliquots of the cell c-ulture supernatants were removed and assayed for levels of IL-2, IL-4, IL-5, IL-6, TNF-alpha and IFN-gamma.
IL-2 (pg/mL) | IL-4 (pg/mL) | IL-5 (pg/mL) | IL-6 (pg/mL) | TNF-alpha (pg/mL) | IFN-gamma (pg/mL) | |
Unstimulated (9 days) | ND | ND | ND | 3.9 | 2.4 | ND |
Stimulated (9 days) | 101,437.7 | 11.5 | 30.9 | 134.6 | 4,519.9 | 39,981.9 |
ND*=Non-detectable
Cited in Article as
kmf00005, SpeedyPLEX Human/Non-Human Primate Th1/Th2 Cytokine (6-plex) CBA Assay Panel, Proteintech, IL, USA