| Test principle: |
This assay employs a two-site sandwich ELISA to quantitate D-lactic acid in samples. An antibody specific for D-lactic acid has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyD-lactic acid present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for D-lactic acid is added to the wells. After washing, Streptavidin conjugated Horseradish Peroxidase (HRP) is added to the wells. Following a wash to remove any unbound avidin-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of D-lactic acid bound in the initial step. The color development is stopped and the intensity of the color is measured.
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| Product Overview: |
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| Components: |
Reagents |
Quantity |
Reagents |
Quantity |
Assay plate (96 Wells) |
1 |
Instruction manual |
1 |
Standard (lyophilized) |
2 |
Sample Diluent |
1 x 20 mL |
Biotin-Conjugate (concentrate 100 x) |
1 x 120 μL |
Biotin-Conjugate Diluent |
1 x 20 mL |
Streptavidin-HRP (concentrate 100 x) |
1 x 120 μL |
Streptavidin-HRP Diluent |
1 x 20 mL |
Wash Buffer (concentrate 25 x) |
1 x 20 mL |
Substrate Solution |
1 x 12 mL |
Stop Solution |
1 x 10 mL |
Adhesive Films |
4 |
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| Specificity: |
This assay has high sensitivity and excellent specificity for detection of Rat D-lactic acid. No significant cross-reactivity or interference between Rat D-lactic acid and analogues was observed.
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| Recovery: |
Matrices listed below were spiked with certain level of recombinant Rat D-lactic acid and the recovery rates were calculated by comparing the measured value to the expected amount of Rat D-lactic acid in samples.
Sample Type |
Number |
Recovery range (%) |
Average(%) |
Serum |
10 |
90-101 |
96 |
EDTA plasma |
10 |
89-97 |
93 |
Heparin plasma |
10 |
91-99 |
95 |
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| Precision: |
Intra-assay Precision (Precision within an assay)
Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.
Inter-assay Precision (Precision between assays)
Three samples of known concentration were tested in forty separate assays to assess inter-assay precision.
CV (%) = SD/meanX100
Intra-Assay: CV<8%
Inter-Assay: CV<12%
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| Linearity: |
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Rat D-lactic acid and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Sample Type |
1：2 |
1：4 |
1：8 |
1：16 |
Serum |
78-89% |
81-99% |
92-103% |
95-105% |
EDTA plasma |
91-101% |
90-98% |
93-101% |
91-98% |
Heparin plasma |
92-103% |
93-102% |
92-99% |
91-101% |
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| Stability: |
The 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.
The loss rate was determined by accelerated thermal degradation test. Keep the kit at 37°C for 4 and 7 days, and compare O.D.values of the kit kept at 37°C with that of at recommended temperature. (referring from China Biological Products Standard, which was calculated by the Arrhenius equation. For ELISA kit, 4 days storage at 37°C can be considered as 6 months at 2 - 8°C, which means 7 days at 37°C equaling 12 months at 2 - 8°C).
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| Sample collection and storage: |
S e r u m : U s e a s e r u m s e p a r a t o r t u b e ( S S T ) a n d a l l o w s a m p l e s t o c l o t f o r t w o h o u r s a t r o o m t e m p e r a t u r e o r o v e r n i g h t a t 2 - 8° C b e f o r e c e n t r i f u g a t i o n f o r 1 5 m i n u t e s a t 1 0 0 0 × g . R e m o v e s e r u m a n d a s s a y i m m e d i a t e l y o r a l i q u o t a n d s t o r e s a m p l e s a t ≤ - 2 0° C . A v o i d r e p e a t e d f r e e z e - t h a w c y c l e s .
P l a s m a : C o l l e c t p l a s m a u s i n g E D T A , o r h e p a r i n a s a n a n t i c o a g u l a n t . C e n t r i f u g e f o r 1 5 m i n u t e s a t 1 0 0 0 × g a t 2 - 8° C w i t h i n 3 0 m i n u t e s o f c o l l e c t i o n . A s s a y i m m e d i a t e l y o r a l i q u o t a n d s t o r e s a m p l e s a t ≤ - 2 0° C . A v o i d r e p e a t e d f r e e z e - t h a w c y c l e s .
O t h e r b i o l o g i c a l f l u i d s : C e n t r i f u g e s a m p l e s f o r 2 0 m i n u t e s a t 1 0 0 0 × g . R e m o v e p a r t i c u l a t e s a n d a s s a y i m m e d i a t e l y o r s t o r e s a m p l e s i n a l i q u o t a t - 2 0° C o r - 8 0° C . A v o i d r e p e a t e d f r e e z e / t h a w c y c l e s .
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| Kits storage instructions: |
Store at 2-8°C. Please refer to Instruction Manual.
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Rat D-lactic acid ELISA Kit
