Nori Sheep IL-12 ELISA Kit
$461.00 – $832.00
DataSheet Â
This ELISA kit is for quantification of IL-12 p35 in sheep. This is a quick ELISA assay that reduces time to 50% compared to the conventional method, and the entire assay only takes 3 hours. This assay employs the quantitative sandwich enzyme immunoassay technique and uses biotin-streptavidin chemistry to improve the performance of the assays. An antibody specific for IL-12 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any IL-12 present is bound by the immobilized antibody. After washing away any unbound substances, a detection antibody specific for IL-12 is added to the wells. Following wash to remove any unbound antibody reagent, a detection reagent is added. After intensive wash a substrate solution is added to the wells and color develops in proportion to the amount of IL-12 bound in the initial step. The color development is stopped, and the intensity of the color is measured.
Alternative names for IL-12: Interleukin 12 P35, IL12 P35
This product is for Laboratory Research Use Only not for diagnostic and therapeutic purposes or any other purposes.
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Description
Nori Sheep IL-12Â ELISA Kit Summary
Alternative names for IL-12: Interleukin 12 P35, IL12 P35
Alternative names for sheep: ovine, lamb, goat
| Assay Type | Solid Phase Sandwich ELISA |
| Format | 96-well Microplate or 96-Well Strip Microplate |
| Method of Detection | Colorimetric |
| Number of Targets Detected | 1 |
| Target Antigen Accession Number | Q9TU27 |
| Assay Length | 3 hours |
| Quantitative/Semiquantitative | Quantitative |
| Sample Type | Plasma, Serum, Cell Culture, Urine, Cell/Tissue Lysates, Synovial Fluid, BAL, |
| Recommended Sample Dilution (Plasma/Serum) | No dilution for sample <ULOQ; sufficient dilution for samples >ULOQ |
| Sensitivity | 12 pg/mL |
| Detection Range | 62.5-4000 pg/mL |
| Specificity | Sheep IL-12 p35 |
| Cross-Reactivity | < 0.5% cross-reactivity observed with available related molecules, < 50% cross-species reactivity observed with species tested. |
| Interference | No significant interference observed with available related molecules |
| Storage/Stability | 4 ºC for up to 6 months |
| Usage | For Laboratory Research Use Only. Not for diagnostic or therapeutic use. |
| Additional Notes | The kit allows for use in multiple experiments. |
Standard Curve
Kit Components
1. Pre-coated 96-well Microplate
2. Biotinylated Detection Antibody
3. Streptavidin-HRP Conjugate
4. Lyophilized Standards
5. TMB One-Step Substrate
6. Stop Solution
7. 20 x PBS
8. Assay Buffer
Other Materials Required but not Provided:
1. Microplate Reader capable of measuring absorption at 450 nm
2. Log-log graph paper or computer and software for ELISA data analysis
3. Precision pipettes (1-1000 µl)
4. Multi-channel pipettes (300 µl)
5. Distilled or deionized water
Protocol Outline
1. Prepare all reagents, samples and standards as instructed in the datasheet.
2. Add 100 µl of Standard or samples to each well and incubate 1 h at RT.
3. Add 100 µl of Working Detection Antibody to each well and incubate 1 h at RT.
4. Add 100 µl of Working Streptavidin-HRP to each well and incubate 20 min at RT.
5. Add 100 µl of Substrate to each well and incubate 5-30 min at RT.
6. Add 50 µl of Stop Solution to each well and read at 450 nm immediately.
Background:Â
Interleukin 12 (IL12) is the founding member of the IL12 family of heterodimeric cytokines, which have important immunological functions. IL12 is composed of two disulfide-linked subunits of 35 kDa and 40 kDa, respectively. The 35 kDa subunit (p35) is an α helical protein homologous to IL6 and GCSF.The 40 kDa subunit (p40) contains one fibronectin type III and one Ig C2-like domain, and has a high degree of structural homology to type I cytokine receptors. Whereas p35 subunit is unique to IL-12, the p40 subunit is also utilized in IL23. Mature rat p40 contains 313 aa and can exist in multiple forms, including monomer, homodimer, heterodimer linked to p19 (forming IL 23), and heterodimer linked to p35 (forming IL12). The expression of p40 is upregulated by substances such as LPS and CpG that activate antigen presenting cells.
Cells known to produce IL12 include macrophages, dendritic cells, monocytes, Langerhans cells, neutrophils, and keratinocytes. The activities of IL12 are mediated by the receptor complex composed of two type I transmembrane proteins: a 100 kDa ligand binding subunit (IL12 Rβ1) and a 130 kDa signal transducing subunit (IL12 Rβ2). IL-12 mediates enhancement of the cytotoxic activity of NK cells and CD8+ cytotoxic T lymphocytes and potentiates the expansion and late activation of Th1 CD4+ T cells. IL-12 also has anti-angiogenic activity (1-4). IL-12 is linked with autoimmunity, and normally stops allergies to food developing (5).
References
- Park, A.Y. and P. Scott (2001) Scand. J. Immunol. 53:529.
- Trinchieri, G. et al. (2003) Immunity 19:641.
- Brombacher, F. et al. (2003) Trends Immunol. 24:207.
- Lankford, C.S. and D.M. Frucht (2003) J. Leukoc. Biol. 73:49
- Temblay JN, et al. J Allergy Clin Immunol. 2007 Sep;120 (3):
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