Eotaxin ELISA Kit (Human) (OKBB00138)

Catalog No: OKBB00138

Size:96 Tests

Human CCL11 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 Eotaxin ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for Eotaxin has been precoated onto 96-well plates. Standards(E.coli, G24-P97) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for Eotaxin 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 Eotaxin 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)	83.4	205	468	84.5	202	485
  St. Dev.	3.5	12.5	19.2	8.2	19.8	44.1
  %CV	4.19	6.09	4.10	9.70	9.80	9.09

• ELISA Kit Component:
  • Lyophilized recombinant human Eotaxin standard: 10 ng/tube x 2
  • Anti-human Eotaxin Antibody Well Plate: 96 Wells
  • Sample diluent buffer: 30 mL
  • Biotinylated anti-human Eotaxin 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 Eotaxin ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. Human Eotaxin 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 Eotaxin 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: Eotaxin, also known as CCL11, is a potent inducer of eosinophil chemotaxis and is considered as a selective ligand of the CC chemokine receptor 3 (CCR3), which is expressed on eosinophils, basophils, and Th2 lymphocytes.1 Eotaxin is assumed to be involved in eosinophilic inflammatory diseases such as atopic dermatitis, allergic rhinitis, asthma and parasitic infections.2 The gene maps to chromosome 17 and is expressed constitutively at high levels in small intestine and colon, and at lower levels in various other tissues. The deduced mature protein sequence is 66% identical to human monocyte chemoattractant protein-1, and 60% identical to guinea pig eotaxin. Recombinant human eotaxin produced in insect cells induces a calcium flux response in normal human eosinophils, but not in neutrophils or monocytes.3 The human eotaxin gene is cloned and found to be 61.8 and 63.2% identical at the amino acid level to guinea pig and mouse eotaxin.4
• 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 (citrate, EDTA)
• Sensitivity: <2 pg/ml
• Predicted Homology Based on Immunogen Sequence: No detectable cross-reactivity with cytokines such as MCP-1, MCP-2, MCP-3 and so on.
• Reference: 1. Ogilvie, P.; Bardi, G.; Clark-Lewis, I.; Baggiolini, M.; Uguccioni, M. Eotaxin is a natural antagonist for CCR2 and an
  agonist for CCR5. Blood 97: 1920-1924, 2001.
  2. Hein, H.; Schluter, C.; Kulke, R.; Christophers, E.; Schroder, J.-M.; Bartels, J. Genomic organization, sequence, and
  transcriptional regulation of the human eotaxin gene. Biochem. Biophys. Res. Commun. 237: 537-542, 1997.
  3. Kitaura, M.; Nakajima, T.; Imai, T.; Harada, S.; Combadiere, C.; Tiffany, H. L.; Murphy, P. M.; Yoshie, O. Molecular
  cloning of human eotaxin, an eosinophil-selective CC chemokine, and identification of a specific eosinophil eotaxin
  receptor, CC chemokine receptor 3. J. Biol. Chem. 271: 7725-7730, 1996.
  4. Ponath, P. D.; Qin, S.; Ringler, D. J.; Clark-Lewis, I.; Wang, J.; Kassam, N.; Smith, H.; Shi, X.; Gonzalo, J.-A.;
  Newman, W.; Gutierrez-Ramos, J.-C.; Mackay, C. R. Cloning of the human eosinophil chemoattractant, eotaxin:
  expression, receptor binding, and functional properties suggest a mechanism for the selective recruitment of
  eosinophils. J. Clin. Invest. 97: 604-612, 1996.
• Specificity: Natural and recombinant human Eotaxin
• Immunogen: Expression system for standard: E.coli; Immunogen sequence: G24-P97
• Description: For quantitative detection of human Eotaxin in cell culture supernatants, serum and plasma(citrate, EDTA).

Target Info

• Gene Symbol: CCL11
• Gene Full Name: chemokine (C-C motif) ligand 11
• Alias Symbols: C-C motif chemokine 11, CCL 11, CCL11, CCL11_HUMAN, Chemokine (C-C motif) ligand 11, Chemokine C C motif ligand 11, Chemokine CC Motif Ligand 11, Chemokine ligand 11, Eosinophil Chemotactic P, Eosinophil chemotactic protein, Eotaxin, Eotaxin 1, MGC22554, SCYA 11, SCYA11, Small inducible cytokine A11, Small inducible cytokine subfamily A (Cys Cys) member 11, Small inducible cytokine subfamily A member 11, Small-inducible cytokine A11
• NCBI Gene Id: 6356
• Protein Name: Eotaxin
• Description of Target: Eotaxin, also known as CCL11, is a potent inducer of eosinophil chemotaxis and is considered as a selective ligand of the CC chemokine receptor 3 (CCR3), which is expressed on eosinophils, basophils, and Th2 lymphocytes.1 Eotaxin is assumed to be involved in eosinophilic inflammatory diseases such as atopic dermatitis, allergic rhinitis, asthma and parasitic infections.2 The gene maps to chromosome 17 and is expressed constitutively at high levels in small intestine and colon, and at lower levels in various other tissues. The deduced mature protein sequence is 66% identical to human monocyte chemoattractant protein-1, and 60% identical to guinea pig eotaxin. Recombinant human eotaxin produced in insect cells induces a calcium flux response in normal human eosinophils, but not in neutrophils or monocytes.3 The human eotaxin gene is cloned and found to be 61.8 and 63.2% identical at the amino acid level to guinea pig and mouse eotaxin.4
• Uniprot ID: P51671
• Protein Accession #: NP_002977.1
• Nucleotide Accession #: NM_002986.2