E-Selectin ELISA Kit (Human) (OKBB00247)

Catalog No: OKBB00247 (Formerly GWB-ZZD042)

Size:96 Tests

Human SELE 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 sE-Selectin ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for sE-Selectin has been precoated onto 96-well plates. Standards(NS0, W22-P556) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for sE-Selectin 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 sE-Selectin amount of sample captured in plate.
• ELISA Kit Range: 125 pg/ml - 8,000 pg/ml
• ELISA Kit Reproducibility:

  	Intra-Assay Precision			Inter-Assay Precision
  Sample	1	2	3	1	2	3
  Mean (ng/mL)	1.7	3.1	4.5	1.6	3.2	4.5
  St. Dev.	0.1	0.16	0.31	0.13	0.21	0.33
  %CV	5.88	5.16	6.88	8.12	6.56	7.33

• ELISA Kit Component:
  • Lyophilized recombinant human E-Selectin standard: 10 ng/tube x 2
  • Anti-human E-Selectin Antibody Well Plate: 96 Wells
  • Sample diluent buffer: 30 mL
  • Biotinylated anti-human E-Selectin 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: 125pg/ml-8000pg/ml
  Principle: Aviva’s human E-Selectin ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. Human E-Selectin specific-specific polyclonal antibodies were precoated onto 96-well plates. The human specific detection monoclonal 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 E-Selectin 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: E-selectin, also called endothelial-leukocyte adhesion molecule-1 (ELAM-1), is a cell surface glycoprotein expressed by cytokine-activated endothelium that mediates the adhesion of blood neutrophils.1 An increased expression of E-selectin has been observed in the arterial endothelium interacting with lymphocytes and macrophages in human atherosclerotic lesions.2 E-selectin plays a critical role in mediating tissue-specific homing of T cells into skin, and of primitive hematopoietic progenitor cells (HPCs) into bone marrow (BM).3 A structurally and functionally related group of genes, lymph node homing receptor (LHR), granule membrane protein 140 (GMP-140), and endothelial leukocyte adhesion molecule 1 (ELAM-1) are shown to constitute a gene cluster on mouse and human chromosome 1.4 The standard product used in this kit is recombinant human E-Selectin, excluding intercellular E-Selectin and transmembrane domain. It consists of totally 535 amino acids with the molecular mass of 58.6KDa.
• 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,citrate)
• Sensitivity: <4 pg/ml
• Predicted Homology Based on Immunogen Sequence: No detectable cross-reactivity with any other cytokine.
• Reference: 1. Collins, T.; Williams, A.; Johnston, G. I.; Kim, J.; Eddy, R.; Shows, T.; Gimbrone, M. A., Jr.; Bevilacqua, M. P.
  Structure and chromosomal location of the gene for endothelial-leukocyte adhesion molecule 1. J. Biol. Chem. 266:
  2466-2473, 1991.
  2. Zheng, F.; Chevalier, J. A.; Zhang, L. Q.; Virgil, D.; Ye, S. Q.; Kwiterovich, P. O. An HphI polymorphism in the
  E-selectin gene is associated with premature coronary artery disease. Clin. Genet. 59: 58-64, 2001.
  3. Dimitroff, CJ.; Lee, JY.; Rafii, S.; Fuhlbrigge, RC.; Sackstein, R. CD44 is a major E-selectin ligand on human
  hematopoietic progenitor cells. J Cell Biol. 2001 Jun 11;153(6):1277-86.
  4. Watson, M. L.; Kingsmore, S. F.; Johnston, G. I.; Siegelman, M. H.; Le Beau, M. M.; Lemons, R. S.; Bora, N. S.;
  Howard, T. A.; Weissman, I. L.; McEver, R. P.; Seldin, M. F. Genomic organization of the selectin family of leukocyte
  adhesion molecules on human and mouse chromosome 1. J. Exp. Med. 172: 263-272, 1990.
• Specificity: Natural and recombinant human sE-Selectin
• Immunogen: Expression system for standard: NS0; Immunogen sequence: W22-P556
• Description: For quantitative detection of human soluble E-Selectin in cell culture supernatants, serum and plasma(heparin, citrate).

Target Info

• Gene Symbol: SELE
• Gene Full Name: selectin E
• Alias Symbols: CD62 antigen-like family member E, CD62E, CD62E antigen, E selectin, E-selectin, ELAM, ELAM-1, ELAM1, Endothelial leukocyte adhesion molecule 1, ESEL, LECAM2, Leukocyte endothelial cell adhesion molecule 2, Leukocyte-endothelial cell adhesion molecule 2, LYAM2_HUMAN, sele
• NCBI Gene Id: 6401
• Protein Name: E-selectin
• Description of Target: E-selectin, also called endothelial-leukocyte adhesion molecule-1 (ELAM-1), is a cell surface glycoprotein expressed by cytokine-activated endothelium that mediates the adhesion of blood neutrophils.1 An increased expression of E-selectin has been observed in the arterial endothelium interacting with lymphocytes and macrophages in human atherosclerotic lesions.2 E-selectin plays a critical role in mediating tissue-specific homing of T cells into skin, and of primitive hematopoietic progenitor cells (HPCs) into bone marrow (BM).3 A structurally and functionally related group of genes, lymph node homing receptor (LHR), granule membrane protein 140 (GMP-140), and endothelial leukocyte adhesion molecule 1 (ELAM-1) are shown to constitute a gene cluster on mouse and human chromosome 1.4 The standard product used in this kit is recombinant human sE-Selectin, excluding intercellular sE-Selectin and transmembrane domain. It consists of totally 535 amino acids with the molecular mass of 58.6KDa.
• Uniprot ID: P16581
• Protein Accession #: NP_000441.2
• Nucleotide Accession #: NM_000450.2