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This kit is based on Double antibody-Sandwich ELISA detection method and takes 4h assay time. The microplate provided in this kit has been precoated with anti IL-12 antibody.
Add standard and properly diluted sample into relevant well respectively.
After incubation, wash unbound components. Add biotinylated detection antibody.
Then, it binds with IL-12 bound to precoated antibody. Wash unbound components and add HRP-Streptavidin Conjugate (SABC).
Wash unbound components again and add TMB substrate solution. Then, TMB was catalyzed by HRP to produce a blue color product that turned yellow after adding a stop solution.
Read the O.D.
absorbance at 450nm in a microplate reader.
Calculate the concentration of IL-12 in the sample by plotting standard curve. The concentration of the target substance is proportional to the OD450 value.
Catalogue No.: EH0176
Alias: IL-12 ELISA Kit, Interleukin-12 subunit alpha ELISA Kit, IL-12A ELISA Kit, Cytotoxic lymphocyte maturation factor 35 kDa subunit ELISA Kit, CLMF p35 ELISA Kit, IL-12 subunit p35 ELISA Kit, NK cell stimulatory factor chain 1 ELISA Kit, NKSF1 ELISA Kit, IL12A ELISA Kit, NKSF1 ELISA Kit, Interleukin-12 subunit beta ELISA Kit, IL-12B ELISA Kit, Cytotoxic lymphocyte maturation factor 40 kDa subunit ELISA Kit, CLMF p40 ELISA Kit, IL-12 subunit p40 ELISA Kit, NK cell stimulatory factor chain 2 ELISA Kit, NKSF2 ELISA Kit, IL12B ELISA Kit, NKSF2 ELISA Kit
Species: Human
Range: 15.625-1000pg/ml
Sensitivity: 9.375pg/ml
Product NameHuman IL-12(Interleukin 12) ELISA KitAliasIL-12 ELISA Kit, Interleukin-12 subunit alpha ELISA Kit, IL-12A ELISA Kit, Cytotoxic lymphocyte maturation factor 35 kDa subunit ELISA Kit, CLMF p35 ELISA Kit, IL-12 subunit p35 ELISA Kit, NK cell stimulatory factor chain 1 ELISA Kit, NKSF1 ELISA Kit, IL12A ELISA Kit, NKSF1 ELISA Kit, Interleukin-12 subunit beta ELISA Kit, IL-12B ELISA Kit, Cytotoxic lymphocyte maturation factor 40 kDa subunit ELISA Kit, CLMF p40 ELISA Kit, IL-12 subunit p40 ELISA Kit, NK cell stimulatory factor chain 2 ELISA Kit, NKSF2 ELISA Kit, IL12B ELISA Kit, NKSF2 ELISA KitCatalogue No.EH0176Size48T/96TSpeciesHumanUniProt No.
P29460 P29459Sample TypeSerum, Plasma, Cell Culture Supernatant, cell or tissue lysate, Other liquid samplesDetection MethodSandwich ELISA, Double AntibodyDetection WavelengthOD450Reaction Duration4 hoursRange15.625-1000pg/mlSensitivity9.
375pg/mlStorage2-8°C(Sealed), Don't cryopreserve.SpecificitySpecifically binds with IL-12 , no obvious cross reaction with other analogues.
ELISA Kit Components
Kit Components — Item — Size(48T) — Size(96T) — Storage Condition for Opened Kit
E001 — ELISA Microplate(Dismountable) — 8×6 — 8×12 — Put the rest strips into a sealed foil bag with the desiccant. Stored for 1 month at 2-8°C; Stored for 6 month at -20°C
E002 — Lyophilized Standard — 1vial — 2vial — Put the rest standards into a desiccant bag. Stored for 1 month at 2-8°C; Stored for 6 month at -20°C
E003 — Biotin-labeled Antibody(Concentrated, 100X) — 60ul — 120ul — 2-8°C (Avoid Direct Light)
E034 — HRP-Streptavidin Conjugate(SABC, 100X) — 60ul — 120ul
E024 — TMB Substrate — 5ml — 10ml
E039 — Sample Dilution Buffer — 10ml — 20ml — 2-8°C
E040 — Antibody Dilution Buffer — 5ml — 10ml
E049 — SABC Dilution Buffer — 5ml — 10ml
E026 — Stop Solution — 5ml — 10ml
E038 — Wash Buffer(Concentrated, 25X) — 15ml — 30ml
E006 — Plate Sealer — 3 pieces — 5 pieces
E007 — Product Description — 1 copy — 1 copy
Required Instruments and Reagents
Assay Procedure Summary
Standard CurveThis product is detected by QC department and meets performance required in the manual. (Laboratory Humidity: 20%-60%; Temperature: 18°C -25°C; Equilibrate TMB substrate to 37°C before staining.
After adding into the ELISA wells, incubate for 15min at 37°C in dark.)
Due to different assay environments and operations, assay data below and standard curve are provided for reference. Experimenters should establish standard curve according to their own assay.
STD.(pg/ml) — OD-1 — OD-2 — Average
0 — 0.093 — 0.098 — 0.095
15.625 — 0.153 — 0.159 — 0.156
31.25 — 0.206 — 0.214 — 0.21
62.5 — 0.29 — 0.302 — 0.296
125 — 0.473 — 0.493 — 0.483
250 — 0.77 — 0.802 — 0.786
500 — 1.397 — 1.454 — 1.425
1000 — 2.095 — 2.181 — 2.138
RecoveryAdd a certain amount of IL-12 into the sample. Calculate the recovery by comparing the measured value with the expected amount of IL-12 in the sample.
Sample Type — Recovery Range(%) — Average(%)
serum(n=10) — 85-104 — 95
EDTA plasma(n=10) — 87-103 — 93
Heparin plasma(n=10) — 86-102 — 94
LinearityDilute the sample with a certain amount of IL-12 at 1:2, 1:4 and 1:8 to get the recovery range.
Sample Type — 1:2 — 1:4 — 1:8
serum(n=10) — 88-104% — 93-103% — 85-104%
EDTA plasma(n=10) — 82-101% — 83-98% — 82-97%
Heparin plasma(n=10) — 83-99% — 81-100% — 82-97%
Precision(%)Intra-assay Precision: samples with low, medium and high concentration are tested 20 times on the same plate.
Inter-assay Precision: samples with low, medium and high concentration are tested 20 times on three different plates.
Item — Intra-assay Precision — Inter-assay Precision
Sample — 1 — 2 — 3 — 1 — 2 — 3
n — 20 — 20 — 20 — 20 — 20 — 20
Mean (pg/ml) — 31.48 — 130.4 — 511 — 32.01 — 123.2 — 502
Standard deviation — 1.69 — 7.55 — 29.23 — 1.71 — 6.96 — 26.56
CV(%) — 5.36 — 5.79 — 5.72 — 5.33 — 5.65 — 5.29
StabilityPerform the stability test for the sealed kit at 37°C and 2-8°C and get relevant data.
ELISA kit(n=5) — 37°C for 1 month — 2-8°C for 6 months
Average(%) — 80 — 95-100
