Laminin ELISA Kit (Human) (OKBB00203)

Catalog No: OKBB00203 (Formerly GWB-CA00J7)

Size:96 Tests

Human LAMININ ELISA Kit standard curve

Product Info

• Predicted Species Reactivity: Human
• Application: ELISA
• ELISA Kit Detection Method: Colorimetric, OD450 nm
• ELISA Kit Duration: ~ 3 Hours
• ELISA Kit Principle: Aviva's human LAMA1 ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A polyclonal antibody from rabbit specific for LAMA1 has been precoated onto 96-well plates. Standards(from human fibroblasts) and test samples are added to the wells, a biotinylated detection polyclonal antibody from rabbit specific for LAMA1 is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The absorbance of yellow is proportional to the human LAMA1 amount of sample captured in plate.
• ELISA Kit Range: 156 pg/ml - 10,000 pg/ml
• ELISA Kit Reproducibility:

  	Intra-Assay Precision			Inter-Assay Precision
  Sample	1	2	3	1	2	3
  Mean (pg/mL)	1335	4334	7066	1728	4566	7324
  St. Dev.	72.09	286	296.8	110.6	342.5	366.2
  %CV	5.4	6.59	4.20	6.40	7.50	5

• ELISA Kit Component:
  • Lyophilized recombinant human Laminin standard: 10 ng/tube x 2
  • Anti-human Laminin Antibody Well Plate: 96 Wells
  • Sample diluent buffer: 30 mL
  • Biotinylated anti-human Laminin antibody: 130 uL, dilution 1:100
  • Antibody diluent buffer: 12 mL
  • Avidin-Biotin-Peroxidase Complex (ABC): 130 uL, dilution 1:100
  • ABC diluent buffer: 12 mL
  • TMB color developing agent: 10 mL
  • TMB stop solution: 10 mL
• Additional Information: Range: 156pg/ml-10,000pg/ml
  Principle: Aviva’s human Laminin ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A polyclonal antibody from rabbit specific for Laminin has been precoated onto 96-well plates. Standards (from human fibroblasts) and test samples are added to the wells, a biotinylated detection polyclonal antibody from rabbit specific for Laminin is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the human Laminin amount of sample captured in plate.
  Notes: 1. To inspect the validity of experiment operation and the appropriateness of sample dilution proportion, pilot
  experiment using standards and a small number of samples is recommended.
  2. The TMB Color Developing agent is colorless and transparent before using, contact us freely if it is not the case.
  3. Before using the Kit, spin tubes and bring down all components to the bottom of tubes.
  4. Duplicate well assay is recommended for both standard and sample testing.
  5. Don’t let 96-well plate dry, for dry plate will inactivate active components on plate.
  6. Don’t reuse tips and tubes to avoid cross contamination.
  7. To avoid to use the reagents from different batches together.
  8. In order to avoid marginal effect of plate incubation due to temperature difference ( reaction may be stronger in the
  marginal wells), it is suggested that the diluted ABC and TMB solution will be pre-warmed in 37C for 30 min before
  using.
  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.
• Reconstitution and Storage: Store at 4C for 6 months, at -20C for 12 months. Avoid multiple freeze-thaw cycles (Shipped with wet ice.)
• Sample Type: cell culture supernates, serum and plasma (heparin, EDTA)
• Sensitivity: <10 pg/ml
• Predicted Homology Based on Immunogen Sequence: No detectable cross-reactivity with any other cytokine
• 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.
• Specificity: Natural human LAMA1
• Immunogen: Expression system for standard: from human fibroblasts
• Description: For quantitative detection of human LAMA1 in cell culture supernatants, cell lysates and serum.

Target Info

• Gene Symbol: LAMA1
• Gene Full Name: laminin, alpha 1; laminin, alpha 2; laminin, beta 1; laminin, beta 2(laminin S)
• Alias Symbols: Lamb2, LAMB2_HUMAN, LAMC1, LAMC2, LAMC3, laminin A, Laminin B, Laminin B1s chain, Laminin gamma 1, Laminin gamma 2, Laminin gamma 3, Laminin subunit beta-2, Laminin-11 subunit beta, Laminin-14 subunit beta, Laminin-15 subunit beta, Laminin-3 subunit beta, Laminin-4 subunit beta, Laminin-7 subunit beta, Laminin-9 subunit beta, OCCM, S-LAM beta, S-laminin subunit beta
• NCBI Gene Id: 3909
• Protein Name: Laminin subunit alpha-1; Laminin subunit alpha-2; Laminin subunit beta-1; Laminin subunit beta-2
• Description of Target: 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.
• Uniprot ID: B0YJ33