For Research Use Only in the United States.
Not for use in diagnostic procedures.
MicroVue Serum PYD provides a quantitative measure of the excretion of pyridinoline cross-link (Pyd) in serum. Pyridinoline cross-links are an indicator of type I collagen resorption, especially bone collagen.
In humans, the total pool of urinary Pyd is approximately 45% free while the remaining fraction is bound to oligopeptides ranging from small linear peptides to very large cross-linked structures in excess of 10,000 Da. The proportion of free to total cross-links appears to be constant in healthy individuals and those with metabolic bone diseases or arthritis conditions, thus providing the rationale for measuring free Pyd.
Method: Competitive
Analyte: Pyridinoline
Specimen: Serum
Specimen Volume: 25 μL post-filtration
LOD*: 0.4 nmol/L
LLOQ*: 0.75 nmol/L
HLOQ*: 12 nmol/L
Precision CVs within-run: 6-15%
Precision CVs between-run: 9-12%
Incubation Time: 20-26 Hours
Cross-reactivity: Free PYD 100% Free DPD 0% PYD/DPD peptides < 2.5% L-arginine 0.0004% (see Package Insert)
*LOD = Limit of Detection, LLOQ = Lower Limit of Quantification, HLOQ = Higher Limit of Quantification
96 well microtiter plate, with reagents sufficient to test 40 samples in duplicate
Free pyridinoline cross-links, some cross-reactivity to glutamine and arginine (0.5 nmol/L signal)
Baboon, Cat, Cow, Dog, Guinea Pig, Horse, Human, Rhesus Macaque, Mouse, Rat
No reactivity detected with Rabbit
Spin serum in Spinfilters (included) for 30 minutes
Add 50 μL Reagent 1 to plate
Add 25 μL serum, diluted Standards, diluted Controls
Add 75 μL cold Pyridinoline Antibody
Incubate overnight at 2°C to 8°C
Wash 3 times with 1x Wash Buffer
Add 150 μL Enzyme Conjugate
Incubate 1 hour at room temperature
Wash 3 times with 1x Wash Buffer
Add 150 μL working Substrate Solution
Incubate 40 +/– 5 minutes at room temperature
Add 100 μL Stop Solution
Read at 405 nm
Data Reduction: 4 Parameter