IL12 ELISA Kit

Catalog No. ABIN1112722

Human IL12 ELISA Kit Colorimetric assay for quantification of Human IL12.

Quick Overview for IL12 ELISA Kit (ABIN1112722)

Target: IL12 (Interleukin 12 (IL12))

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 7.8-500 pg/mL

Application: ELISA

Product Details IL12 ELISA Kit

Minimum Detection Limit: 7.8 pg/mL

Analytical Method: Quantitative

Sensitivity: < 2 pg/mL

Components: 1. One 96-well plate pre-coated with anti-Human IL-12(P70) antibody 2. Lyophilized Human IL-12(P70) standards: 2 tubes (10ng / tube) 3. Sample / Standard diluent buffer: 30ml 4. Biotin conjugated anti-Human IL-12(P70) antibody (Concentrated): 130 µl.

Material not included: 1. 37 °C incubator 2. Microplate reader (wavelength: 450nm) 3. Precise pipette and disposable pipette tips 4. Automated plate washer 5. ELISA shaker 6. 1.5ml of Eppendorf tubes 7. Plate cover 8. Absorbent filter papers 9. Plastic or glass container with volume of above 1L

Application Details

Comment:

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti-IL-12(P70) polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti-IL-12(P70) polyclonal antibody was used as detection antibodies. The standards test samples and biotin conjugated detection antibody were added - the wells subsequently and wash with wash buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with wash buffer. TMB substrates were used - visualize HRP enzymatic reaction. TMB was catalyzed by HRP - produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional - the IL-12(P70) amount of sample captured in plate. Read the O.D. absorbance at 450 nm in a microplate reader and then the concentration of IL-12(P70) can be calculated.

Plate: Pre-coated

Reagent Preparation:

1. Before the experiment, centrifuge each kit component for several minutes to bring down all reagents to the bottom of tubes. 2. It is recommend to measure each standard and sample in duplicate. 3. Do NOT let the plate completely dry at any time! Since the dry condition can inactivate the biological material on the plate. 4. Do not reuse pipette tips and tubes to avoid cross contamination. 5. Do not use the expired components and the components from different batches. 6. To avoid the marginal effect of plate incubation for temperature differences (the marginal wells always get stronger reaction), it is recommend to equilibrate the ABC working solution and TMB substrate for at least 30 min at room temperature (37°C ) before adding to wells.The TMB substrate (Kit Component 8) is colorless and transparent before use, if not, please contact us for replacement.

Sample Preparation:

Preparation of sample and reagents 1. Sample Isolate the test samples soon after collecting, then, analyze immediately (within 2 hours). Or aliquot and store at -20 °C for long term. Avoid multiple freeze-thaw cycles.
Cell culture supernatants, tissue lysate or body fluids: Centrifuge to remove precipitate, analyze immediately or aliquot and store at -20 °C° C.
Serum: Coagulate the serum at room temperature (about 30 min). Centrifuge at approximately 1000 × g for 10 min. Analyze the serum immediately or aliquot and store at -20 °C . Note: 1. Coagulate blood samples completely, then, centrifuge, and avoid hemolysis and particle. 2. NaN3 can not be used as test sample preservative, since it is the inhibitor for HRP.

Restrictions: For Research Use only

Handling

Preservative: Sodium azide, Thimerosal (Merthiolate)

Target Details for IL12

Target: IL12 (Interleukin 12 (IL12))

Alternative Name: IL-12 P70

Background: Interleukin 12 (IL-12) is an interleukin that is naturally produced by dendritic cells, macrophages and human B-lymphoblastoid cells (NC-37) in response to antigenic stimulation. IL-12 is composed of a bundle of four alpha helices. It is a heterodimeric cytokine encoded by two separate genes, IL-12A (p35) and IL-12B (p40). IL-12 plays an important role in the activities of natural killer cells and T lymphocytes. It mediates enhancement of the cytotoxic activity of NK cells and CD8+ cytotoxic T lymphocytes. IL-12 also has anti-angiogenic activity, which means it can block the formation of new blood vessels.

Pathways: JAK-STAT Signaling, TLR Signaling, Cellular Response to Molecule of Bacterial Origin, Regulation of Leukocyte Mediated Immunity, Positive Regulation of Immune Effector Process, Activated T Cell Proliferation, Cancer Immune Checkpoints, Inflammasome