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This assay recognizes recombinant and natural rat Kim-1.

Mouse procollagen III propeptide (P&IIIP) ELISA Kit


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Background

Different kinds of collagen have been identified in human. All of them derive from longer precursor molecules (procollagens). They are synthesized intracellularly and secreted in extracellular space where they are cleaved by aminoproteases. Among the different kinds of precursors, type III is one of the most abundant interstitial procollagens. Since its aminoterminal propeptide, PIIIP, is formed in equimolar proportions to collagen, serum measurements of this fragment can provide an index of collagen synthesis.

Feature and Advantages

<PRINCIPLE OF THE ASSAY>
The microtiter plate provided in this kit has been pre-coated with an antibody specific to P?P. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated polyclonal antibody preparation specific for P?P and Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. Then a TMB (3,3'5, 5' tetramethyl-benzidine) substrate solution is added to each well. Only those wells that contain P?P, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of a sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The concentration of P?P in the samples is then determined by comparing the O.D. of the samples to the standard curve.

< SPECIFICITY>
This assay recognizes recombinant and natural mouse P?P. No significant cross-reactivity or interference was observed.

<DETECTION RANGE>
0.78 ng/ml-50 ng/ml
The standard curve concentrations used for the ELISA’s were 50 ng/ml, 25 ng/ml, 12.5 ng/ml, 6.25 ng/ml, 3.12 ng/ml, 1.56 ng/ml, 0.78 ng/ml.

<SENSITIVITY>
The minimum detectable dose of mouse P?P is typically less than 1.95 ng/ml. The sensitivity of this assay, or Lower Limit of Detection (LLD) was defined as the lowest protein concentration that could be differentiated from zero.

Kit component


Reagent Quantity
1. Assay plate 1
2. Standard 2
3. Sample Diluent 1 x 20 ml
4. Biotin-antibody Diluent 1 x 10 ml
5. HRP-avidin Diluent 1 x 10 ml
6. Biotin-antibodyt 1 x 120μl
7. HRP-avidin 1 x 120μl
8. Wash Buffer 1 x 20 ml (25×concentrate)
9. TMB Substrate 1 x 10 ml
10. Stop Solution 1 x 10 ml
Product List

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Species : 0.78 ng/ml - 50 ng/ml
Assay range : 100 µL (Plasma, Serum)
Assay sample volume : 6-8 hr.{3 hr


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