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View additional product information for IL-12 p70 Rat Singleplex Kit - FAQs (LRC0121)
4 product FAQs found
Reduction of the sample volume by half, 25 uL, is a viable alternative when sample volume is limiting.
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The following is a list of general lab supplies that are required for running BioSource immunoassays on the Luminex xMAP system:
1) Sonicating water bath
2) Orbital shaker
3) Vortexer
4) Repeating and/or multi-channel pipetter (not required, but recommended)
5) Calibrated adjustable precision pipettes, with disposable plastic tips
6) Glass/plastic tubes and racks for preparing reagents
7) Graduated cylinder and container for preparing wash solution
8) Aluminum foil
9) Deionized or distilled water.
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The Luminex beads should be protected from light because they are susceptible to photobleaching. We recommend protecting the beads by keeping containers covered with aluminum foil during all incubation steps, and exercising care during handling. The beads should not be frozen, subjected to excessive heat, or exposed to organic solvents.
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The capture and the detection antibody in our Luminex kits LHC0121 (human specific), LMC0121 (mouse specific), and LRC0121 (rat specific) bind to different epitopes on the p40 subunit. These kits therefore can measure the free monomeric p40 subunit, the p40-p40 homodimer, and the p40-p35 heterodimer (designated p70). It is this p40-p35 heterodimer (i.e., p70) which is the active form of the IL-12. The p40 gene and the p35 gene bear no homology to one another and are located on different chromosomes. In humans, the p40 gene is located in the 5p31-q33 region, while the p35 gene is located in the 3p12-p13.2 region. Transcription of these two genes is differentially regulated. While the p35 gene is expressed by all cell types, the expression of the p40 gene is highly regulated, containing at least four transcription factor-binding sites, and limited only to IL-12 producing cells. Interestingly, monomeric p40 is found to be secreted in large excess over the p70 heterodimer (D'Andrea et al., 1992), and p40-p40 homodimers are believed to play a role in antagonizing the biological activity of the p70 active form. Kang et al., 1996 found that epidermal Langerhans cells, when cultured and allowed to mature into dendritic cells produce both p40 and p70. D'Andrea et al., 1992 found that macrophages stimulated with Staphaylococcus aureus or LPS also produce both p40 and p70. Other cell types that have been observed to transcribe both IL-12 genes include bone marrow cells cultured in the presence of stem cell factor, keratincytes, and EBV transformed B cells.
References:
Kang, K., M. Kubin, K.D. Cooper, S.R. Lessin, G. Trinchieri, and A.H. Rook (1996) IL-12 synthesis by human Langerhans cells. J. Immunol. 156:1402-1407.
D'Andrea, A., M. Rengaraju, N.M. Valiante, J. Chehimi, M. Kubin, M. Aste, S.H. Chan, M. Kobayashi, D. Young, E. Nickbarg, R. Chizzonite, S.F. Wolf, and G. Trinchieri (1992) Production of a natural killer cell stimulatory factor (interleukin-12) by peripheral blood mononuclear cells. J. Exp. Med. 176:1387-1398.
Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.