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ELISA Kit for Platelet Activating Factor (PAF)

PAF-Acether; AGEPC; Acetyl-Glyceryl-Ether-Phosphorylcholine

Specificity

This assay has high sensitivity and excellent specificity for detection of Platelet Activating Factor (PAF).
No significant cross-reactivity or interference between Platelet Activating Factor (PAF) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Platelet Activating Factor (PAF) and the recovery rates were calculated by comparing the measured value to the expected amount of Platelet Activating Factor (PAF) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 79-93 83
EDTA plasma(n=5) 98-105 102
heparin plasma(n=5) 95-103 98

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Platelet Activating Factor (PAF) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Platelet Activating Factor (PAF) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Platelet Activating Factor (PAF) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 78-105% 85-101% 98-105% 91-101%
EDTA plasma(n=5) 92-103% 81-102% 98-105% 97-105%
heparin plasma(n=5) 98-105% 92-101% 89-103% 78-102%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 50µL standard or sample to each well.
    And then add 50µL prepared Detection Reagent A immediately.
    Shake and mix. Incubate 1 hour at 37°C;
3. Aspirate and wash 3 times;
4. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
5. Aspirate and wash 5 times;
6. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
7. Add 50µL Stop Solution. Read at 450 nm immediately.

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Magazine Citations
PLOS ONE Lysophosphatidylcholine Acyltransferase1 Overexpression Promotes Oral Squamous Cell Carcinoma Progression via Enhanced Biosynthesis of Platelet-Activating Factor Pubmed:25803864
J Biol Chem. Platelet-activating factor contributes to Bacillus anthracis lethal toxin-associated damage. Pubmed:24478317
Mediators of Inflammation Mediators of Mast Cells in Bullous Pemphigoid and Dermatitis Herpetiformis Pubmed:25400334
Molecular Medicine Reports Comparing the role of Ginkgolide B and Ginkgolide K on cultured astrocytes exposed to oxygen‑glucose deprivation Pubmed: 30221704
BioMed Research International Chlorogenic Acid Attenuates Dextran Sodium Sulfate-Induced Ulcerative Colitis in Mice through MAPK/ERK/JNK Pathway
Contraception Antifertility effectiveness of a novel polymer matrix composite and its influence on the endometrium in rhesus macaques (Macaca mulatta) Pubmed: 30980831
Catalog No. Related products for research use of Pan-species (General) Organism species Applications (RESEARCH USE ONLY!)
CEA526Ge ELISA Kit for Platelet Activating Factor (PAF) Enzyme-linked immunosorbent assay for Antigen Detection.
LMA526Ge Multiplex Assay Kit for Platelet Activating Factor (PAF) ,etc. by FLIA (Flow Luminescence Immunoassay) FLIA Kit for Antigen Detection.