MicroVue tDPD EIA**

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.

• Allows quantitative measure of Dpd in serum
• Results in urine are comparable to HPLC

Features & Benefits

Feature	Benefit

Product Specifications

Format	ELISA
Test/kit	96 wells/plate
Sample type	Serum or urine

Performance Characteristics

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

About the AssayFormat

96-well microplate with reagents sufficient to test 39 samples in duplicate

Antibody Specificity

Free Dpd (all Dpd is hydrolyzed to free form); <1% reactivity with pyridinoline

Species Reactivity

Baboon, Cow, Dog, Guinea Pig, Horse, Mouse, Rat, Rabbit, Squirrel Monkey, Cynomolgus Monkey, Rhesus Macaque, Human Cell Culture, Pig, Sheep, Cat

Hydrolysis Steps

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

Assay Steps

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

Ordering Information

Catalog Number	Description
8032	96 Wells/Plate

Product Documentation

Package Insert

SDS

Technical Data Sheet

Technical Bulletin - ELISA Wash Technique