General Hyaluronic Acid (HA) ELISA Kit
two product lines: Traditional ELISA Kit and Ready-to-Use ELISA Kit.
The kit is a competitive inhibition enzyme immunoassay technique for the in vitro quantitative measurement of HA in general serum, plasma or other biological fluids.
|Composition||Traditional ELISA Kit||Ready-to-Use ELISA KIT||Conform|
|Pre-coated, ready to use 96-well strip plate 1||Pre-coated, ready to use 96-well strip plate 1|
|Plate sealer for 96 wells 2||Plate sealer for 96 wells 2|
|Standard 2||Standard 2|
|Diluents buffer 1×45mL||Standard Diluent 1×20mL|
|Detection Reagent A 1×120μL||Detection Solution A 1×12mL|
|Detection Reagent B 1×120μL||Detection Solution B 1×12mL|
|TMB Substrate 1×9mL||TMB Substrate 1×9mL|
|Stop Solution 1×6mL||Stop Solution 1×6mL|
|Wash Buffer (30 × concentrate) 1×20mL||Wash Buffer (30 × concentrate) 1×20mL|
|Instruction manual 1||Instruction manual 1|
This assay employs the competitive inhibition enzyme immunoassay technique. A monoclonal antibody specific to the index has been pre-coated onto a microplate. A competitive inhibition reaction is launched between biotin labeled the index and unlabeled the index (Standards or samples) with the pre-coated antibody specific to the index. After incubation the unbound conjugate is washed off. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. The amount of bound HRP conjugate is reverse proportional to the concentration of the index in the sample. After addition of the substrate solution, the intensity of color developed is reverse proportional to the concentration of the index in the sample.
Matrices listed below were spiked with certain level of recombinant the index and the recovery rates were calculated by comparing the measured value to the expected amount of the index in samples.
|Matrix||Recovery range (%)||Average(%)|
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of the index and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
PrecisionIntra-assay Precision (Precision within an assay): 3 samples with low, middle and high level the index were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level the index were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
StabilityThe stability of ELISA 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 unnecessary influences on the performance, operation procedures and lab conditions, especially room temperature, air humidity, and incubator temperatures should be strictly regulated. It is also strongly suggested that the whole assay is performed by the same experimenter from the beginning to the end.
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℃;
3. Aspirate and wash 3 times;
4. Add 100µL prepared Detection Reagent B. Incubate 1 hour at 37℃;
5. Aspirate and wash 5 times;
6. Add 90µL Substrate Solution. Incubate 15-25 minutes at 37℃;
7. Add 50µL Stop Solution. Read at 450 nm immediately.
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