For Research Use Only. Not for Use in Diagnostic Procedures
The MicroVue tDPD kit provides reagents for hydrolysis of specimens and measurement of total Dpd in urine and serum.
Reagents and controls for assaying total deoxypyridinoline (Dpd) crosslinks in serum or urine.
In humans, the total pool of urinary Dpd 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. Free and total cross-links appear in healthy individuals and those with metabolic bone diseases, thus providing the rationale for measuring the combined total forms of Dpd. Improvements in immunoassay methodology have resulted in the ability to measure Total Dpd level in serum and urine thus permitting a novel method for researching bone specific collagen degradation.
ELISA
96 wells/plate
Serum or urine
Method: Competitive Analyte: Deoxypyridinoline Specimen: Serum or Urine Specimen Volume: 100 µL Serum, 50 µL Urine LOD*: 0.5 nmol/L LLOQ*: 1 nmol/L HLOQ*: 100 nmol/L Precision CVs within-run: 5-12% Precision CVs between-run: 10-17% Incubation Time: 23-25 Hours Cross-reactivity: Free Dpd 100% Free Pyd < 1% Pyd/Dpd peptides < 2.5%
*LOD = Limit of Detection, LLOQ = Lower Limit of Quantification, HLOQ = Higher Limit of Quantification
96-well microplate with reagents sufficient to test 39 samples in duplicate
Free Dpd (all Dpd is hydrolyzed to free form); <1% reactivity with pyridinoline
Baboon, Cow, Dog, Guinea Pig, Horse, Mouse, Rat, Rabbit, Squirrel Monkey, Cynomolgus Monkey, Rhesus Macaque, Human Cell Culture, Pig, Sheep, Cat
For serum samples Mix samples and Serum Hydrolysis Control with acid reagent Centrifuge for 5 minutes Add supernatants and Serum Hydrolysis Control to hydrolysis plate
For urine samples Dilute samples and Urine Hydrolysis Control with water Add acid reagent to hydrolysis plates Add diluted samples and reconstituted Urine Hydrolysis Control to hydrolysis plate Incubate 18-20 hours at 99°C Cool to 4°C Neutralize with base reagent
Add assay Buffer to all wells Add diluted Standards and Controls and hydrolyzed/neutralized samples and Hydrolysis Control Incubate 30 minutes at 2°C to 8°C Add Enzyme Conjugate Incubate 2 hours at 2°C to 8°C Wash 3 times with Wash Buffer Add Substrate Incubate 2 hours at room temperature Add Stop Solution Read at 405 nm Data Reduction: 4 Parameter Correct urinary results for creatinine concentration
96 Wells/Plate
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