Cat Deoxypyridinoline (DPD) ELISA Kit

This assay employs a two-site sandwich ELISA to quantitate DPD in samples. An antibody specific for DPD has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any DPD present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for DPD is added to the wells. After washing, Streptavidin conjugated Horseradish Peroxidase (HRP) is added to the wells. Following a wash to remove any unbound avidin-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of DPD bound in the initial step. The color development is stopped and the intensity of the color is measured.
 

Deoxypyridinoline (Dpd) is one of the two pyridinium cross-links that provide structural rigidity to type I collagen in bone. During osteoclastic resorption, Dpd is released into circulation and is excreted in the urine in free and peptide-bound forms.Deoxypyridinoline (Dpd) occurs mainly in Type I collagen of bone. After the formation of the Type I collagen matrix, the action of lysyl oxidase on two hydroxylysine and a lysine residue result in the formation of a Dpd cross link between two triple helical strands of collagen. Deoxypyridinoline (Dpd) has been described as a marker of bone turnover in metastatic breast cancer.Pyridinoline (PYD) and Deoxypyridinoline (DPD) are the major stable cross-links. These cross-links, primarily in bone and cartilage, have been found in all the connective tissues except the skin. PYD is the major component of these tissues, whereas DPD has only been detected in bone and dentine.
 

Reagents	Quantity	Reagents	Quantity
Assay plate (96 Wells)	1	Instruction manual	1
Standard (lyophilized)	2	Sample Diluent	1 x 20 mL
Biotin-Conjugate (concentrate 100 x)	1 x 120 μL	Biotin-Conjugate Diluent	1 x 12 mL
Streptavidin-HRP  (concentrate 100 x)	1 x 120 μL	Streptavidin-HRP Diluent	1 x 12 mL
Wash Buffer (concentrate 25 x)	1 x 20 mL	Substrate Solution	1 x 10 mL
Stop Solution	1 x 6 mL	Adhesive Films	4

 

This assay has high sensitivity and excellent specificity for detection of Human DPD. No significant cross-reactivity or interference between Human DPD and analogues was observed.
 

Matrices listed below were spiked with certain level of recombinant Human DPD and the recovery rates were calculated by comparing the measured value to the expected amount of Human DPD in samples.

Sample Type	Number	Recovery range (%)	Average(%)
Serum	10	90-101	96
EDTA plasma	10	89-97	93
Heparin plasma	10	91-99	95

 

Intra-assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision. Inter-assay Precision (Precision between assays) Three samples of known concentration were tested in forty separate assays to assess inter-assay precision. CV (%) = SD/meanX100 Intra-Assay: CV<8% Inter-Assay: CV<12%
 

Intra-assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision. Inter-assay Precision (Precision between assays) Three samples of known concentration were tested in forty separate assays to assess inter-assay precision. CV (%) = SD/meanX100 Intra-Assay: CV<8% Inter-Assay: CV<12%

Sample Type	1：2	1：4	1：8	1：16
Serum	78-89%	81-99%	92-103%	95-105%
EDTA plasma	91-101%	90-98%	93-101%	91-98%
Heparin plasma	92-103%	93-102%	92-99%	91-101%

 

Intra-assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision. Inter-assay Precision (Precision between assays) Three samples of known concentration were tested in forty separate assays to assess inter-assay precision. CV (%) = SD/meanX100 Intra-Assay: CV<8% Inter-Assay: CV<12%
 

 

Store at 2-8°C. Please refer to Instruction Manual.