Human CD31/PECAM-1 Quantikine ELISA Kit

Catalog # Availability Size / Price Qty
DCD310
Control Products Available
Human CD31/PECAM-1 ELISA Cell Culture Supernate/Saliva/Urine Standard Curve
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Human CD31/PECAM-1 Quantikine ELISA Kit Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 uL), Serum (50 uL), EDTA Plasma (50 uL), Heparin Plasma (50 uL), Saliva (50 uL), Urine (50 uL)
Sensitivity
0.078 ng/mL
Assay Range
0.3 - 20 ng/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Saliva, Urine)
Specificity
Natural and recombinant human CD31.
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
Interference
No significant interference observed with available related molecules.

Product Summary

The Quantikine Human CD31/PECAM-1 Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human CD31 in cell culture supernates, serum, plasma, saliva, and urine. It contains CHO cell-expressed recombinant human CD31 and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human CD31 showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for naturally occurring human CD31.

Precision

Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in twenty separate assays to assess inter-assay precision. Assays were performed by at least three technicians using two lots of components

Cell Culture Supernates, Saliva, Urine

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean 1.62 4.7 9.96 1.71 5 10.6
Standard Deviation 0.065 0.117 0.372 0.104 0.252 0.411
CV% 4 2.5 3.7 6.1 5 3.9

Serum, EDTA Plasma, Heparin Plasma

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean 2.01 5.86 12.3 1.9 5.71 12
Standard Deviation 0.07 0.182 0.403 0.163 0.363 0.677
CV% 3.5 3.1 3.3 8.6 6.4 5.6

Recovery

The recovery of human CD31 spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 103 96-112
EDTA Plasma (n=4) 100 97-106
Heparin Plasma (n=4) 101 94-108
Serum (n=4) 104 100-107

Linearity

To assess the linearity of the assay, samples containing and/or spiked with high concentrations of human CD31 were diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human CD31/PECAM-1 ELISA Linearity

Product Datasheets

Preparation and Storage

Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: CD31/PECAM-1

CD31/PECAM-1 is a heavily glycosylated transmembrane homophilic adhesion protein that is highly expressed on endothelial cells and at a lower level on platelets, granulocytes, macrophages, dendritic cells, T and B cells, and NK cells. It is required for the transepithelial migration of leukocytes. CD31 signaling increases the activation threshold of T cells and inhibits dendritic cell development. The extracellular domain of CD31 is released during endothelial cell apoptosis. This fragment circulates in the serum of patients suffering from myocardial infarction, acute ischaemic stroke, and multiple sclerosis, conditions that involve tissue damage and endothelial cell apoptosis.

Long Name:
Platelet Endothelial Cell Adhesion Molecule 1
Entrez Gene IDs:
5175 (Human); 18613 (Mouse); 29583 (Rat)
Alternate Names:
adhesion molecule; CD31 antigen; CD31; CD31/EndoCAM; endoCAM; FLJ34100; FLJ58394; GPIIA'; PECA1; PECAM1; PECAM-1; PECAM-1, CD31/EndoCAM; platelet endothelial cell adhesion molecule; platelet/endothelial cell adhesion molecule
⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.

Assay Procedure

Refer to the product for complete assay procedure.

Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
  1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
  2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

  3. 100 µL Assay Diluent
  4.   Add 100 µL of Assay Diluent to each well.

  5. 50 µL Standard, Control, or Sample
  6.   Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours on a horizontal orbital microplate shaker.
  7.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

  8. 200 µL Conjugate
  9.   Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours on the shaker.
  10.   Aspirate and wash 4 times.

  11. 200 µL Substrate Solution
  12.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.

  13. 50 µL Stop Solution
  14.   Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.

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