MCP-1 ELISA Kit (Human) (OKBB00210)

Catalog No: OKBB00210

Size:96 Tests

Human MCP-1 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 MCP-1 ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A polyclonal antibody from goat specific for MCP-1 has been precoated onto 96-well plates. Standards(E.coli, 24-T99) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for MCP-1 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 MCP-1 amount of sample captured in plate.
• ELISA Kit Range: 15.6 pg/ml - 1,000 pg/ml
• ELISA Kit Reproducibility:

  	Intra-Assay Precision			Inter-Assay Precision
  Sample	1	2	3	1	2	3
  Mean (pg/mL)	73	362	663	93	397	695
  St. Dev.	2.7	17.74	34.5	5.022	24.22	51.43
  %CV	3.69	4.90	5.20	5.4	6.10	7.4

• ELISA Kit Component:
  • Lyophilized recombinant human IL-12(p40) standard: 10 ng/tube x 2
  • Anti-human IL-12(p40) Antibody Well Plate: 96 Wells
  • Sample diluent buffer: 30 mL
  • Biotinylated anti-human IL-12(p40) 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: 15.6pg/ml-1000pg/ml
  Principle: Aviva’s human MCP-1 ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. Human MCP-1 specific-specific polyclonal antibodies were precoated onto 96-well plates. The human specific detection polyclonal antibodies were biotinylated. The test samples and biotinylated detection antibodies were added to the wells 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 MCP-1 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: Monocyte chemoattractant protein-1 (MCP-1), a member of the chemokine (chemotactic cytokine) family, is a potent monocyte agonist that is upregulated by oxidized lipids.1 MCP-1 is also known as CCL2, SCYA2, MCAF. MCAF is a member of family of factors involved in immune and inflammatory responses. The amino acid sequence deduced from the nucleotide sequence reveals the primary structure of the MCAF precursor to be composed of a putative signal peptide sequence of 23 amino acid residues and a mature MCAF sequence of 76 amino acid residues. 2 MCP-1 plays a unique and crucial role in the initiation of atherosclerosis and may provide a new therapeutic target in this disorder.3 Human MCP-1 is a 8.7KDa non-glycoprotein, consisting of 99 amino acids in precursor form and 76 amino acids in mature form.
• 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, plasma (heparin, EDTA, citrate) and urine
• Sensitivity: <1 pg/ml
• Predicted Homology Based on Immunogen Sequence: Cross-reactivates with human Eotaxin, MCP-3 <1%.
• Reference: 1. Gosling, J.; Slaymaker, S.; Gu, L.; Tseng, S.; Zlot, C. H.; Young, S. G.; Rollins, B. J.; Charo, I. F. MCP-1 deficiency
  reduces susceptibility to atherosclerosis in mice that overexpress human apolipoprotein B. J. Clin. Invest. 103:
  773-778, 1999.
  2. Furutani, Y.; Nomura, H.; Notake, M.; Oyamada, Y.; Fukui, T.; Yamada, M.; Larsen, C. G.; Oppenheim, J. J.;
  Matsushima, K. Cloning and sequencing of the cDNA for human monocyte chemotactic and activating factor
  (MCAF). Biochem. Biophys. Res. Commun. 159: 249-255, 1989.
  3. Gu, L.; Okaka, Y.; Clinton, S. K.; Gerard, C.; Sukhova, G. K.; Libby, P.; Rollins, B. J. Absence of monocyte
  chemoattractant protein-1 reduces atherosclerosis in low density lipoprotein receptor-deficient mice. Molec. Cell 2:
  275-281, 1998.
• Specificity: Natural and recombinant human MCP-1
• Immunogen: Expression system for standard: E.coli; Immunogen sequence: Q24-T99
• Description: For quantitative detection of human MCP-1 in cell culture supernatants, serum, plasma(heparin, EDTA, citrate) and urine.

Target Info

• Gene Symbol: CCL2
• Gene Full Name: chemokine (C-C motif) ligand 2
• Alias Symbols: C-C motif chemokine 2, CCL2, CCL2_HUMAN, Chemokine (C-C motif) ligand 2, GDCF-2, HC11, HSMCR30, MCAF, MCP-1, MCP1, MGC9434, Monocyte chemoattractant protein 1, Monocyte chemoattractant protein-1, Monocyte chemotactic and activating factor, Monocyte chemotactic protein 1, Monocyte secretory protein JE, SCYA2, Small inducible cytokine A2 (monocyte chemotactic protein 1, homologous to mouse Sig-je), Small inducible cytokine subfamily A (Cys-Cys), member 2, Small-inducible cytokine A2, SMC-CF
• NCBI Gene Id: 6347
• Protein Name: C-C motif chemokine 2
• Description of Target: Monocyte chemoattractant protein-1 (MCP-1), a member of the chemokine (chemotactic cytokine) family, is a potent monocyte agonist that is upregulated by oxidized lipids.1 MCP-1 is also known as CCL2, SCYA2, MCAF. MCAF is a member of family of factors involved in immune and inflammatory responses. The amino acid sequence deduced from the nucleotide sequence reveals the primary structure of the MCAF precursor to be composed of a putative signal peptide sequence of 23 amino acid residues and a mature MCAF sequence of 76 amino acid residues.2 MCP-1 plays a unique and crucial role in the initiation of atherosclerosis and may provide a new therapeutic target in this disorder.3 Human MCP-1 is a 8.7KDa non-glycoprotein, consisting of 99 amino acids in precursor form and 76 amino acids in mature form.
• Uniprot ID: P13500
• Protein Accession #: NP_002973.1
• Nucleotide Accession #: NM_002982.3