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ELISA Kit for High Density Lipoprotein (HDL)

Specificity

This assay has high sensitivity and excellent specificity for detection of High Density Lipoprotein (HDL).
No significant cross-reactivity or interference between High Density Lipoprotein (HDL) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant High Density Lipoprotein (HDL) and the recovery rates were calculated by comparing the measured value to the expected amount of High Density Lipoprotein (HDL) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 80-92 89
EDTA plasma(n=5) 83-93 87
heparin plasma(n=5) 94-102 98

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level High Density Lipoprotein (HDL) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level High Density Lipoprotein (HDL) 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 High Density Lipoprotein (HDL) 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) 89-97% 82-98% 82-104% 88-95%
EDTA plasma(n=5) 81-93% 95-103% 96-103% 80-97%
heparin plasma(n=5) 92-105% 87-95% 97-105% 80-89%

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 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.

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Magazine Citations
Iranian Journal of Health and Physical activity Effects of Aerobic Training, with or without Zizyphus Jujuba Water Extraction, on Fundus Nesfatin-1, ATP, HDL-C, and LDL-C Concentrations in Female Rats Ijvst: Source
Scandinavian Journal of Medicine & Science in Sports Endurance training selectively increases HDL-bound sphingosine-1-phosphate in the plasma pubmed:28493600
Journal of Nutritional Science and Vitaminology (Tokyo) Anthocyanin Cyanidin-3-Glucoside Attenuates Platelet Granule Release in Mice Fed High-Fat Diets. pubmed:28978870
RESPIRATORY RESEARCH The HDL from septic-ARDS patients with composition changes exacerbates pulmonary endothelial dysfunction and acute lung injury induced by cecal ligation … Pubmed: 33148285
respiratory research The adverse alterations of HDL in quality promote septic ARDS via exacerbated pulmonary endothelial dysfunction
Diabetes Metab Syndr Obes ¡°Adjusting Internal Organs and Dredging Channel¡± Electroacupuncture Ameliorates Insulin Resistance in Type 2 Diabetes Mellitus by Regulating the Intestinal?¡­ 34135611
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