Human Homocysteine(HCY)ELISA Kit
Product Details
| Description |
The Human Homocysteine (HCY) ELISA Kit allows for the in vitro quantitative determination of HCY concentrations in serum, plasma, urine, or tissue homogenates. This assay kit was designed and optimized for metabolic research use in humans. It has undergone rigorous quality control in multiple parameters, including sensitivity, specificity, precision, linearity, recovery, and inter-batch difference. Refer to the product instructions for more details. This assay employs the quantitative sandwich enzyme immunoassay technique, in which HCY in the samples or standards are sandwiched between pre-coated HCY antibody and Biotin-conjugated HCY antibody. HRP-avidin is then added into the wells. Following a wash to remove any unbound reagent, the TMB substrate solution is added to the wells and color develops in proportion to the amount of HCY bound in the initial step. The color development is stopped upon adding the stop solution, and the intensity of the color is measured at 450 nm via a micro... Read more |
| Target Name |
Homocysteine(HCY) |
| Alternative Names |
N/A |
| Abbreviation |
Hcy |
| Species |
Homo sapiens (Human) |
| Sample Types |
serum, plasma, urine, tissue homogenates |
| Detection Range |
0.78 nmol/mL-50 nmol/mL |
| Sensitivity |
0.195 nmol/mL |
| Assay Time |
1-5h |
| Sample Volume |
50-100ul |
| Detection Wavelength |
450 nm |
| Research Area |
Metabolism |
| Assay Principle |
quantitative |
| Measurement |
Sandwich |
| Precision |
| Intra-assay Precision (Precision within an assay): CV% |
| Three samples of known concentration were tested twenty times on one plate to assess. |
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| Inter-assay Precision (Precision between assays): CV% |
| Three samples of known concentration were tested in twenty assays to assess. |
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| Linearity |
| To assess the linearity of the assay, samples were spiked with high concentrations of human HCY in various matrices and diluted with the Sample Diluent to produce samples with values within the dynamic range of the assay. |
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Sample |
Serum(n=4) |
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| 1:1 |
Average % |
85 |
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| Range % |
80-90 |
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| 1:2 |
Average % |
98 |
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| Range % |
91-105 |
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| 1:4 |
Average % |
89 |
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| Range % |
84-96 |
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| 1:8 |
Average % |
93 |
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| Range % |
86-98 |
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| Recovery |
| The recovery of human HCY spiked to levels throughout the range of the assay in various matrices was evaluated. Samples were diluted prior to assay as directed in the Sample Preparation section. |
| Sample Type |
Average % Recovery |
Range |
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| Serum (n=5) |
96 |
91-101 |
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| EDTA plasma (n=4) |
92 |
89-94 |
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| Typical Data |
| These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed. |
| nmol/ml |
OD1 |
OD2 |
Average |
Corrected |
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| 50 |
2.082 |
2.156 |
2.119 |
1.997 |
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| 25 |
1.456 |
1.489 |
1.473 |
1.351 |
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| 12.5 |
0.941 |
0.968 |
0.955 |
0.833 |
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| 6.25 |
0.583 |
0.597 |
0.590 |
0.468 |
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| 3.125 |
0.342 |
0.367 |
0.355 |
0.233 |
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| 1.56 |
0.248 |
0.261 |
0.255 |
0.133 |
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| 0.78 |
0.192 |
0.198 |
0.195 |
0.073 |
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| 0 |
0.121 |
0.123 |
0.122 |
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| Materials provided |
- A micro ELISA plate --- The 96-well plate has been pre-coated with an anti-human HCY antibody. This dismountable microplate can be divided into 12 x 8 strip plates.
- Two vials lyophilized standard ---Dilute a bottle of the standard at dilution series, read the OD values, and then draw a standard curve.
- One vial Biotin-labeled HCY antibody (100 x concentrate) (120 μl/bottle) ---Act as the detection antibody.
- One vial HRP-avidin (100 x concentrate) (120 μl/bottle) ---Bind to the detection antibody and react with the TMB substrate to make the solution chromogenic.
- One vial Biotin-antibodyDiluent (15 ml/bottle) ---Dilute the Biotin-antibody.
- One vial HRP-avidin Diluent (15 ml/bottle) ---Dilute the HRP-avidin solution.
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| Materials not provided |
- A microplate reader capable of measuring absorbance at 450 nm, with the correction wavelength set at 540 nm or 570 nm.
- An incubator can provide stable incubation conditions up to 37°C±5°C.
- Centrifuge
- Vortex
- Squirt bottle, manifold dispenser, or automated microplate washer
- Absorbent paper for blotting the microtiter plate
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| Storage |
Store at 2-8°C. Please refer to protocol. |
| Lead Time |
3-5 working days |
