Human IL-12(Interleukin 12) ELISA Kit

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

1. Microplate reader (wavelength: 450nm)
2. 37°C incubator (CO2 incubator for cell culture is not recommenced.)
3. Automated plate washer or multi-channel pipette/5ml pipettor (for manual washing purpose)
4. Precision single (0.5-10μL, 5-50μL, 20-200μL, 200-1000μL) and multi-channel pipette with disposable tips(Calibration is required before use.)
5. Sterile tubes and Eppendorf tubes with disposable tips
6. Absorbent paper and loading slot
7. Deionized or distilled water

Assay Procedure Summary

• Step 1: Add 100ul standard or sample into each well, seal the plate and statically incubate for 90 minutes at 37°C.
• Washing: Wash the plate twice without immersing.
• Step 2: Add 100ul biotin-antibody working solution, seal the plate and statically incubate for 60 minutes at 37°C.
• Washing: Wash the plate three times and immerse for 1min each time.
• Step 3: Add 100ul HRP-Streptavidin Conjugate (SABC) working solution, seal the plate and statically incubate for 30 minutes at 37°C.
• Washing: Wash the plate five times and immerse for 1min each time.
• Step 4: Add 90ul TMB substrate solution, seal the plate and statically incubate for 10-20 minutes at 37°C. (Accurate TMB visualization control is required.)
• Step 5: Add 50ul stop solution. Read at 450nm immediately and calculate.

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.

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