ELISA Kit for Keratan Sulfate (KS)

Keratosulfate

Specificity

This assay has high sensitivity and excellent specificity for detection of Keratan Sulfate (KS).
No significant cross-reactivity or interference between Keratan Sulfate (KS) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Keratan Sulfate (KS) and the recovery rates were calculated by comparing the measured value to the expected amount of Keratan Sulfate (KS) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 90-103 99
EDTA plasma(n=5) 97-105 101
heparin plasma(n=5) 93-101 97

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Keratan Sulfate (KS) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Keratan Sulfate (KS) 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 Keratan Sulfate (KS) 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) 83-99% 95-103% 87-97% 90-99%
EDTA plasma(n=5) 96-104% 79-93% 94-105% 98-105%
heparin plasma(n=5) 80-99% 80-99% 81-93% 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 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
Journal of Clinical Medicine The Effects of TNF-α Inhibition on the Metabolism of Cartilage: Relationship between KS, HA, HAPLN1 and ADAMTS4, ADAMTS5, TOS and TGF-β1 Plasma … Pubmed:35407621
Catalog No. Related products for research use of Homo sapiens (Human) Organism species Applications (RESEARCH USE ONLY!)
CEA590Hu ELISA Kit for Keratan Sulfate (KS) Enzyme-linked immunosorbent assay for Antigen Detection.