Human CD62P ELISA Assay

$635.00

The Human CD62P ELISA Assay is to be used for the in-vitro quantitative determination of CD62P (P-Selectin), in supernatant, buffered solutions or serum and plasma samples. The assay will recognize both natural and recombinant human CD62P. The Eagle Biosciences Human CD62P ELISA Assay kit is for Research Use Only.

SKU: C2P31-K01 Categories: , ,

Human CD62P ELISA Assay

The Human CD62P ELISA Assay is For Research Use Only

Size: 1×96 wells
Sensitivity: 6 ng/ml
Dynamic Range: 6.87 – 220 ng/ml
Incubation Time: Overnight
Sample Type: Serum, Plasma, Cell Culture
Sample Size: 100 µl
Alternative Names: P-selectin, LECAM-3


Summary

P-selectin (CD62P, LECAM-3) was originally purified from platelets and later found to be expressed also in endothelial cells. In both cell types, P-selectin is constitutively expressed and stored in secretory granules – α-granules in platelets and Weibel-Palade bodies in endothelial cells.

A soluble form of P-selectin, which might represent a proteolytic fragment or a soluble splice variant lacking the transmembrane domain, is found in serum and plasma. High expression of P-selectin or sP-selectin has been implicated in several inflammatory disorders, including adult respiratory distress syndrome, acute lung injury, ischemia-reperfusion injury, Gram-negative septic shock, thrombotic diseases, malaria, systemic sclerosis, connective tissue disease and rheumatoid arthritis. Furthermore, lung injury scores correlate significantly with sP-selectin in plasma of patients with acute lung injury.


Storage Instructions
Store the CD62P ELISA Assay kit reagents between 2 and 8°C. Immediately after use remaining reagents should be returned to cold storage (2-8°C). Expiry of the kit and reagents is stated on box front labels. The expiry of the kit components can only be guaranteed if the components are stored properly, and if, in case of repeated use of one component, the reagent is not contaminated by the first handling.


Related Products

Human CD62E ELISA Assay
Human CD62L ELISA Assay
Human CD31 ELISA Assay

Additional Information

Assay Principle


The CD62P Kit is a solid phase sandwich Enzyme Linked-Immuno-Sorbent Assay (ELISA). A monoclonal antibody specific for CD62P has been coated onto the wells of the microtiter strips provided. Samples, including standards of known CD62P concentrations and unknowns are pipetted into these wells.

During the first incubation, the CD62P antigen, standards and zeros are bound this is followed by a separate incubation where the biotinylated monoclonal antibody specific for CD62P is added.

After washing, the enzyme (streptavidin-peroxydase) is added. After incubation and washing to remove all the unbound enzyme, a substrate solution is added to induce a coloured reaction product. The intensity of this coloured product is directly proportional to the concentration of CD62P present in the samples.

  1. Add 100µl of each standard, sample and zero (Standard Diluent) in duplicate to appropriate number of wells
  2. Cover with a plastic plate cover and incubate at room temperature (18 to 25°C) for 1 hour
  3. Remove the cover and wash the plate
  4. Add 50µl of diluted biotinylated anti-CD62P to all wells
  5. Cover with a plastic plate cover and incubate at room temperature (18 to 25°C) for 1 hour
  6. Repeat wash step 4.
  7. Add 100μl of Streptavidin-HRP solution into all wells
  8. Cover with a plastic plate cover and incubate at room temperature (18 to 25°C) for 30 min
  9. Repeat wash step 4.
  10. Add 100μl of ready-to-use TMB Substrate Solution into all wells
  11. Incubate in the dark for 10-20 minutes* at room temperature. Avoid direct exposure to light by wrapping the plate in aluminium foil
  12. Add 100μl of H2SO4:Stop Reagent into all wells

Typical Standard Curve


Specificity


Ten specificities were tested in the Human CD62P ELISA Assay Kit. No cross reaction was observed for MICA, gp130, GM-CSF, IL-2, IL-6, TRAIL, IL-13, IL-8, Perforin and CD31.

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