Product Information
Product Name: SpeedyPLEX Human Cytokine (4-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-4 | 7.81-2000 | 0.52 |
| IL-6 | 3.91-1000 | 0.15 |
| IL-1 alpha | 0.78-200 | 0.02 |
| TNF-alpha | 3.91-1000 | 0.62 |
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 4B1 | 30 mL/bottle | 1 bottle | |
Detection Diluent | 30 mL/bottle | 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-4, IL-6, IL-1 alpha and TNF-alpha in this assay, the concentration measured are shown below.
Analyte | Range (pg/mL) | No. of Detectable (n) | Mean (pg/mL) | % of Detectable |
IL-4 | ND | 0 | ND | 0 |
IL-6 | ND-1.67 | 1 | 0.1 | 8 |
IL-1 alpha | ND-0.37 | 1 | 0 | 8 |
TNF-alpha | ND-5.62 | 5 | 1.3 | 42 |
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 uM β-mercaptoethanol, 2 mM L-glutamine, 100 U/mL penicillin, and 100 ug/mL streptomycin sulfate. Cells were cultured unstimulated or stimulated with 10 ug/mL PHA for 3 days and 5 days. Aliquots of the Cell culture supernatants were removed a-nd assayed for levels of human IL-4, human IL-6, human IL-1 alpha and human TNF-alpha.
Condition | IL-4 (pg/mL) | IL-6 (pg/mL) | IL-1 alpha (pg/mL) | TNF-alpha (pg/mL) |
Unstimulated (3 days) | ND | 59.4 | 42.0 | 16.0 |
Stimulated (3 days) | ND | 21782.9 | 223.8 | 6126.8 |
Unstimulated (5 days) | ND | 8.0 | 16.7 | 34.0 |
Stimulated (5 days) | ND | 29101.0 | 433.3 | 6935.8 |
Human peripheral blood mononuclear cells (0.5 x 106 cells/mL) were cultured in RPMI supplemented with 10% fetal bovine serum, 5 ug/mL PHA and 10 ng/mL recombinant human IL-2. After 7 d-ays of culture, cells were changed into DMEM with 10% fetal bovine serum, 10 ng/mL PMA, 1 ug/mL Ca2+ ionophore and 5 ug/mL PHA and cultured for 1 day. An aliquot of the culture supernatant-s was removed and assayed for human IL-4, human IL-6, human IL-1 alpha and human TNF-alpha.
Condition | IL-4 (pg/mL) | IL-6 (pg/mL) | IL-1 alpha (pg/mL) | TNF-alpha (pg/mL) |
Unstimulated | 17.0 | 10.5 | 11.7 | 443.0 |
Stimulated | 331.6 | 54.9 | 50 | 24520.8 |
ND*=Non-detectable
Cited in Article as
kmf00001, SpeedyPLEX Human Cytokine (4-plex) CBA Assay Panel, Proteintech, IL, USA