This Human Interleukin 12 (IL-12/P70) ELISA Kit is suitable for the quantitative detection of IL-12 concentrations in human serum, cell culture supernates, or tissue homogenates. This assay employs the sandwich ELISA technique in which IL-12 in the sample is sandwiched between the pre-coated anti-human IL-12 antibody and Biotin-labeled IL-12 antibody and the Ag-Ab-Ag complex is labeled by the HRP-avidin and then develops color reaction after the addition of the TMB substrate solution. The color intensity can be measured by a microplate reader at 450 nm. The kit has been validated with high sensitivity, excellent specificity, precision low than 10%, high recovery, and consistency between batches.
IL-12 is a pro-inflammatory cytokine that belongs to the IL-12 family. As a heterodimeric protein, IL-12 consists of two subunits: p35 and p40. IL-12 is primarily generated by activated antigen-presenting cells, such as dendritic cells and macrophages. It plays a critical role in the promotion of Th1 immune responses by inducing IFN-γ production to fight against pathogens and malignant tumors.
||Homo sapiens (Human)
||serum, cell culture supernates, tissue homogenates
||4.7 pg/mL-300 pg/mL
Intra-assay Precision (Precision within an assay): CV%<8%
Three samples of known concentration were tested twenty times on one plate to assess.
Inter-assay Precision (Precision between assays): CV%<10%
Three samples of known concentration were tested in twenty assays to assess.
||To assess the linearity of the assay, samples were spiked with high concentrations of human IL-12/P70 in various matrices and diluted with the Sample Diluent to produce samples with values within the dynamic range of the assay.
||The recovery of human IL-12/P70 spiked to levels throughout the range of the assay in various matrices was evaluated. Samples were diluted
||These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed.
- A micro ELISA plate ---The 96-well plate has been pre-coated with an anti-human IL-12 antibody. This dismountable microplate can be divided into 12 x 8 strip plates.
- Two vials lyophilized standard ---Dilute a bottle of the standard at dilution series, read the OD values, and then draw a standard curve.
- One vial Biotin-labeled IL-12 antibody (100 x concentrate) (120 μl/bottle) ---Act as the detection antibody.
- One vial HRP-avidin (100 x concentrate) (120 μl/bottle) ---Bind to the detection antibody and react with the TMB substrate to make the solution chromogenic.
- One vial Biotin-antibody Diluent (15 ml/bottle) ---Dilute the Biotin-antibody.
- One vial HRP-avidin Diluent (15 ml/bottle) ---Dilute the HRP-avidin solution.
- One vial Sample Diluent (50 ml/bottle)---Dilute the sample to an appropriate concentration.
- One vial Wash Buffer (25 x concentrate) (20 ml/bottle) ---Wash away unbound or free substances.
- One vial TMB Substrate (10 ml/bottle) ---Act as the chromogenic agent. TMB interacts with HRP, eliciting the solution turns blue.
- One vial Stop Solution (10 ml/bottle) ---Stop the color reaction. The solution color immediately turns from blue to yellow.
- Four Adhesive Strips (For 96 wells) --- Cover the microplate when incubation.
- An instruction manual
|Materials not provided
- A microplate reader capable of measuring absorbance at 450 nm, with the correction wavelength set at 540 nm or 570 nm.
- An incubator can provide stable incubation conditions up to 37°C±5°C.
- Squirt bottle, manifold dispenser, or automated microplate washer
- Absorbent paper for blotting the microtiter plate
- 50-300ul multi-channel micropipette
- Pipette tips
- Single-channel micropipette with different ranges
- 100ml and 500ml graduated cylinders
- Deionized or distilled water
- Test tubes for dilution
|ELISA kit FAQs
||Store at 2-8°C. Please refer to protocol.
||3-5 working days