P-Selectin ELISA Kit (Mouse) (OKBB00251)

Catalog No: OKBB00251

Size:96 Tests

Mouse SELP ELISA Kit standard curve

Product Info

• Predicted Species Reactivity: Mouse
• Application: ELISA
• ELISA Kit Detection Method: Colorimetric, OD450 nm
• ELISA Kit Duration: ~ 3 Hours
• ELISA Kit Principle: Aviva's mouse P-Selectin ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from rat specific for P-Selectin has been precoated onto 96-well plates. Standards(NS0, W42-A709) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for P-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 mouse P-Selectin amount of sample captured in plate.
• ELISA Kit Range: 312 pg/ml - 20,000 pg/ml
• ELISA Kit Reproducibility:

  	Intra-Assay Precision			Inter-Assay Precision
  Sample	1	2	3	1	2	3
  Mean (pg/mL)	325	874	1886	417	1029	2020
  St. Dev.	14	43.7	71.7	27.11	63.8	94.94
  %CV	4.30	5	3.80	6.50	6.20	4.7

• ELISA Kit Component:
  • Lyophilized recombinant mouse P-Selectin standard: 20 ng/tube x 2
  • Anti-mouse P-Selectin Antibody Well Plate: 96 Wells
  • Sample diluent buffer: 30 mL
  • Biotinylated anti-mouse P-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: 62.5pg/ml-4000pg/ml
  Principle: Aviva’s mouse P-Selectin ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from rat specific for P-Selectin has been precoated onto 96-well plates. Standards (NSO, W42-A709) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for P-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 density of yellow is proportional to the mouse P-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: P-selectin, also called GMP-140, CD62, or selectin P, is a 140-kD adhesion molecule, expressed at the surface of activated cells, that mediates the interaction of activated endothelial cells or platelets with leukocytes.1 It is stored in secretory granules and expressed at the plasma membrane after cell activation.2 It is known to play an important role in atherosclerosis.3 The major ligand for P-selectin on leukocytes is P-selectin glycoprotein ligand-1 (PSGL-1).4 The standard product used in this kit is recombinant mouse P-Selectin, excluding intercellular P-Selectin and transmembrane domain. It has 42-709 amino acids sequence with the molecular mass of 99.1KDa. As a result of glycosylation, the molecular mass of 190-200KDa is revealed by SDS-PAGE
• 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)
• Sensitivity: <5 pg/ml
• Predicted Homology Based on Immunogen Sequence: No detectable cross-reactivity with any other cytokine.
• Reference: 1. Herrmann, S.-M.; Ricard, S.; Nicaud, V.; Mallet, C.; Evans, A.; Ruidavets, J.-B.; Arveiler, D.; Luc, G.; Cambien, F.
  The P-selectin gene is highly polymorphic: reduced frequency of the pro715 allele carriers in patients with
  myocardial infarction. Hum. Molec. Genet. 7: 1277-1284, 1998.
  2. Florian, V.; Schluter, T.; Bohnensack, R. A new member of the sorting nexin family interacts with the C-terminus of
  P-selectin. Biochem. Biophys. Res. Commun. 281: 1045-1050, 2001.
  3. Burger, P. C.; Wagner, D. D. Platelet P-selectin facilitates atherosclerotic lesion development. Blood 101: 2661-2666,
  2003.
  4. Aigner, S.; Sthoeger, ZM.; Fogel, M.; Weber E.; Zarn, J.; Ruppert, M.; Zeller, Y.; Vestweber, D.; Stahel, R.; Sammar,
  M.; Altevogt, P. CD24, a mucin-type glycoprotein, is a ligand for P-selectin on human tumor cells. Blood. 1997 May
  1;89(9):3385-95.
• Specificity: Natural and recombinant mouse sP-Selectin
• Immunogen: Expression system for standard: NS0; Immunogen sequence: W42-A709
• Description: For quantitative detection of mouse P-Selectin in cell culture supernatants, serum and plasma (citrate).

Target Info

• Gene Symbol: SELP
• Gene Full Name: selectin P (granule membrane protein 140kDa, antigen CD62)
• Alias Symbols: Antigen CD62, CD62, CD62 antigen like family member P, CD62 antigen-like family member P, CD62P, FLJ45155, GMP 140, GMP-140, GMP140, Granule membrane protein 140, Granule membrane protein 140kDa, Granulocyte membrane protein, GRMP, LECAM3, Leukocyte endothelial cell adhesion molecule 3, Leukocyte-endothelial cell adhesion molecule 3, LYAM3_HUMAN, P selectin, P-selectin, PADGEM, Platelet activation dependent granule external membrane protein, Platelet activation dependent granule-external membrane protein, Platelet alpha granule membrane protein, PSEL, Selectin P, Selectin P (granule membrane protein 140kDa antigen CD62), SELP
• NCBI Gene Id: 20344
• Protein Name: P-selectin
• Description of Target: P-selectin, also called GMP-140, CD62, or selectin P, is a 140-kD adhesion molecule, expressed at the surface of activated cells, that mediates the interaction of activated endothelial cells or platelets with leukocytes.1 It is stored in secretory granules and expressed at the plasma membrane after cell activation.2 It is known to play an important role in atherosclerosis.3 The major ligand for P-selectin on leukocytes is P-selectin glycoprotein ligand-1 (PSGL-1).4 The standard product used in this kit is recombinant mouse P-Selectin, excluding intercellular P-Selectin and transmembrane domain. It has 42-709 amino acids sequence with the molecular mass of 99.1KDa. As a result of glycosylation, the molecular mass of 190-200KDa is revealed by SDS-PAGE.
• Uniprot ID: Q01102
• Protein Accession #: NP_035477.1
• Nucleotide Accession #: NM_011347.2