ELISA Kit for Homocysteine (HCy)

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 86-96 90
EDTA plasma(n=5) 93-102 97
heparin plasma(n=5) 93-102 99

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Homocysteine (HCy) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Homocysteine (HCy) 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 Homocysteine (HCy) 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) 95-102% 95-104% 98-105% 84-102%
EDTA plasma(n=5) 80-101% 89-98% 79-89% 97-104%
heparin plasma(n=5) 83-95% 96-103% 87-94% 88-102%

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 Developmental Origins of Health and Disease Maternal low-quality protein diet exerts sex-specific effects on plasma amino acid profile and alters hepatic expression of methyltransferases in adult rat offspring Pubmed:29582727
The Scientific Journal of Al-Azhar Medical Faculty, Girls Homocysteinemia in relation to anemia in hypothyroid patients Doi: 10.4103/sjamf.sjamf_31_18
Science of The Total Environment Organochlorine pesticides exposure may disturb homocysteine metabolism in pregnant women Pubmed: 31787282
AMERICAN JOURNAL OF TRANSLATIONAL RESEARCH Baicalin rescues hyperglycemia-induced neural tube defects via targeting on retinoic acid signaling Pubmed: 32774702
Journal of Diabetes Research Changes and Risk Factors of Skeletal Muscle Mass and Strength in Patients with Type 2 Diabetes over 60 Years Old: A Cross-Sectional Study from China
Journal of Advances in Medical and Pharmaceutical Sciences Homocysteinaemia in Heart Failure Patients in North Eastern Nigeria
Obstetrik ve Neonatoloji T?p Dergisi KARD?YOVASK¨¹LER S?STEM HASTALIKLARINDA R?SK FAKT?R¨¹ OLAN H?PERHOMOS?STE?NEM?'N?N POL?K?ST?K OVER SENDROMUNDAK? YER?
Frontiers in Nutrition Folate, Vitamin B12, and Homocysteine Levels in Women With Neural Tube Defect-Affected Pregnancy in Addis Ababa, Ethiopia Pubmed:35464038
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