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 Technical information: High Sensitive Keratan Sulfate ELISA Kit |
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| - Factors Affecting Measurements (2) |
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[ Recovery Test ]
We used the High Sensitive Keratan Sulfate ELISA Kit to carry out recovery testing for different types of animal serum.
Method
Various animal serum were prepared as shown in the table below, left to stand at room temperature (15-25°C) for 1 h, and measured in accordance with the protocol. The Keratan Sulfate concentrations were calculated and recovery rates derived.
| Addition group (A) | 1 part by volume of sample + 1 part by volume of standard solution |
| Non-addition group (B) | 1 part by volume of sample + 1 part by volume of reaction buffer solution |
| Addition amount (C) | 1 part by volume of reaction buffer solution + 1 part by volume of standard solution |
Results
| Sample | Dilution | Addition
group (A) (ng/mL) |
Non-addition
group (B) (ng/mL) |
Amount recovered (A) - (B) (ng/mL) |
Amount added (C) (ng/mL) |
Recovery rate |
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| Serum | Mouse | 1/4 | 1.92 | 1.24 | 0.68 | 1.12 | 60.7 |
| Rat | 1/40 | 2.10 | 0.91 | 1.19 | 1.11 | 107.2 | |
| Guinea pig | 1/200 | 2.38 | 1.40 | 0.98 | 1.00 | 98.0 | |
| Rabbit | 1/40 | 1.79 | 0.94 | 0.85 | 1.04 | 81.7 | |
| Dog | 1/40 | 1.80 | 0.96 | 0.84 | 1.02 | 82.4 | |
| Human | 1/800 | 2.56 | 1.52 | 1.04 | 1.06 | 98.1 | |
| Recovery rate= | (A) - (B) | X100 |
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| (C) |
Conclusion
Good addition recovery rates of 81.7-107.2% were obtained for rat, guinea pig, rabbit, dog, and human serum. These results verified that the High Sensitive Keratan Sulfate ELISA Kit can be used for measurements irrespective of the effect of other substances also present in serum.
A result of 60.7% was obtained for mouse serum, somewhat lower than the results for other animals, indicating that serum components may exert a slight effect. Enzyme digestion tests of mouse serum, however, exhibited good specificity and dilution linearity, meaning that semiquantitative measurements of Keratan Sulfate in mouse serum are possible.
