MIP-1I ELISA Kit (Human) (OKBB00215)

Catalog No: OKBB00215

Size:96 Tests

Human CCL3 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 MIP-1 alpha ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for MIP-1 alpha has been precoated onto 96-well plates. Standards(E.coli, A27-A92) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for MIP-1 alpha 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 MIP-1 alpha 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)	107	450	732	95	413	682
  St. Dev.	4.39	13.05	25.62	6.84	22.3	34.78
  %CV	4.10	2.9	3.5	7.2	5.39	5.09

• 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: 31.2pg/ml-2000pg/ml
  Principle: Aviva’s human MIP-1α ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. Human MIP-1α specific-specific monoclonal 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 MIP-1α 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: Macrophage inflammatory protein-1 alpha (MIP-1 alpha), also called CCL3, LD78. The cDNA for MIP-1 alpha predicts a mature peptide of 69 amino acids with a molecular mass of 7,889 daltons.1 LD78 is a member of a newly identified superfamily of small inducible proteins involved in inflammatory responses, wound healing, and tumorigenesis.2 MIP-1 alpha is a chemokine that has pro-inflammatory and stem cell inhibitory activities in vitro.3 It constitutes an important second signal for mast cell degranulation in the conjunctiva in vivo and consequently for acute-phase disease.4 The standard product used in this kit is recombinant human MIP-1α, consisting of 66 amino acids with the molecular mass of 7.5kDa.
• 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, citrate)
• Sensitivity: <10 pg/ml
• Predicted Homology Based on Immunogen Sequence: No detectable cross-reactivity with any other cytokine
• Reference: 1. Wolpe, S. D.; Cerami, A. Macrophage inflammatory proteins 1 and 2: members of a novel superfamily of cytokines.
  FASEB J. 3: 2565-2573, 1989.
  2. Nakao, M.; Nomiyama, H.; Shimada, K. Structures of human genes coding for cytokine LD78 and their expression.
  Molec. Cell. Biol. 10: 3646-3658, 1990.
  3. Cook, D. N.; Beck, M. A.; Coffman, T. M.; Kirby, S. L.; Sheridan, J. F.; Pragnell, I. B.; Smithies, O. Requirement of
  MIP-1-alpha for an inflammatory response to viral infection. Science 269: 1583-1585, 1995.
  4. Miyazaki, D.; Nakamura, T.; Toda, M.; Cheung-Chau, K.-W.; Richardson, R. M.; Ono, S. J. Macrophage
  inflammatory protein-1-alpha as a costimulatory signal for mast cell-mediated immediate hypersensitivity reactions.
  J. Clin. Invest. 115: 434-442, 2005.
• Specificity: Natural and recombinant human MIP-1 alpha
• Immunogen: Expression system for standard: E.coli; Immunogen sequence: A27-A92
• Description: For quantitative detection of human MIP-1 alpha in cell culture supernatants, serum and plasma(heparin, EDTA, citrate).

Target Info

• Gene Symbol: CCL3
• Gene Full Name: chemokine (C-C motif) ligand 3
• Alias Symbols: C C motif chemokine 3, CCL3, CCL3_HUMAN, Chemokine (C C motif) ligand 3, Chemokine ligand 3, G0/G1 switch regulatory protein 19 1, G0/G1 switch regulatory protein 19-1, G0S19 1, G0S19 1 protein, Heparin binding chemotaxis protein, L2G25B, LD78-alpha(4-69), LD78ALPHA, Macrophage inflammatory protein 1 alpha, Macrophage inflammatory protein 1-alpha, MIP 1 alpha, MIP-1-alpha, MIP-1-alpha(4-69), MIP1A, PAT 464.1, SCYA 3, SCYA3, SIS alpha, SIS beta, SIS-beta, Small inducible cytokine A3, Small-inducible cytokine A3, Tonsillar lymphocyte LD78 alpha protein
• NCBI Gene Id: 6348
• Protein Name: C-C motif chemokine 3
• Description of Target: Macrophage inflammatory protein-1 alpha (MIP-1 alpha), also called CCL3, LD78. The cDNA for MIP-1 alpha predicts a mature peptide of 69 amino acids with a molecular mass of 7,889 daltons.1 LD78 is a member of a newly identified superfamily of small inducible proteins involved in inflammatory responses, wound healing and tumorigenesis. MIP-1 alpha is a chemokine that has pro-inflammatory and stem cell inhibitory activities in vitro. It constitutes an important second signal for mast cell degranulation in the conjunctiva in vivo and consequently for acute-phase disease. The standard product used in this kit is recombinant human MIP-1α, consisting of 66 amino acids with the molecular mass of 7.5kDa.
• Uniprot ID: P10147
• Protein Accession #: NP_002974.1
• Nucleotide Accession #: NM_002983.2