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EK000017-EK0435: Rat Laminin ELISA Kit
Range 156pg/ml-10,000pg/ml
Sensitivity < 10pg/ml
Specificity no detectable cross-reactivity with any other cytokine.
Application: for quantitative detection of rat Laminin in sera, plasma, body fluids, tissue lysates or cell culture supernatant.
Expiration: four months at 4°C and eight months at -20°C.
Background
Laminin is a large basement membrane glycoprotein composed of three subunits designated the A, B1, and B2.1 Laminin has diverse biological functions, which include stimulating epithelial cell growth and differentiation.2 The nucleotide sequence of human laminin A chain has an open reading frame encoding 3075-amino acids.1 The human laminin A chain is at locus 18p11.3.3 The nucleotide sequence of the human laminin B1 reveals a 5358-base pair open reading frame that potentially codes for 1786 amino acids, including 20 amino acids of a presumptive signal peptide.2 The gene for the human laminin-B1 chain has been localized to chromosome 7, band q31.4 The B2 chain consists of six distinct domains, including two domains with alpha-helical, coiled-coil structures, two domains with cysteine-rich homologous repeats, and two globular domains. The amino acid sequences of the B2 and B1 chains demonstrate considerable homology.5 The human laminin B2 chain gene maps to the long arm of chromosome 1 in the band q31.6 The standard product used in this kit is isolated from plasma.
Principle
The ELISA Kit is based on standard sandwich enzyme-linked immune-sorbent assay technology. The target specific antibodies are precoated onto 96-well plates. The target from the sample is bound to the microwell. The biotinylated target specific detection antibodies are added to the microwells and followed by washing with the PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex is added and unbound conjugates are washed away with the PBS or TBS buffer. TMB, the HRP (horseradish peroxidase) substrate, is used to visualize color change resulting from the HRP enzymatic reaction. TMB is catalyzed by HRP to produce the blue color. The color changes into yellow after the acidic stop solution is added. The density of the yellow color is proportional to the target amount from the sample captured in the microwells.
Reference
1. Haaparanta, T.; Uitto, J.; Ruoslahti, E.; Engvall, E. Molecular cloning of the cDNA encoding human laminin A chain. Matrix 11: 151-160, 1991.
2. Sasaki, M.; Kato, S.; Kohno, K.; Martin, G. R.; Yamada, Y. Sequence of the cDNA encoding the laminin B1 chain reveals a multidomain protein containing cysteine-rich repeats. Proc. Nat. Acad. Sci. 84: 935-939, 1987.
3. Nagayoshi, T.; Mattei, M.-G.; Passage, E.; Knowlton, R.; Chu, M.-L.; Uitto, J. Human laminin A chain (LAMA) gene: chromosomal mapping to locus 18p11.3. Genomics 5: 932-935, 1989.
4. Jaye, M.; Modi, W. S.; Ricca, G. A.; Mudd, R.; Chiu, I.-M.; O'Brien, S. J.; Drohan, W. N. Isolation of a cDNA clone for the human laminin-B1 chain and its gene localization. Am. J. Hum. Genet. 41: 605-615, 1987.
5. Sasaki, M.; Yamada, Y. The laminin B2 chain has a multidomain structure homologous to the B1 chain. J. Biol. Chem. 262: 17111-17117, 1987.
6. Mattei, M.-G.; Weil, D.; Pribula-Conway, D.; Bernard, M. P.; Passage, E.; Van Cong, N.; Timpl, R.; Chu, M.-L. cDNA cloning, expression and mapping of human laminin B2 gene to chromosome 1q31. Hum. Genet. 79: 235-241, 1988.
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