This immunoassay kit allows for the in vitro quantitative determination of human Versican concentrations in cell culture supernates, serum, plasma and other biological fluids.

Human versican/PG-M/PG-350 ELISA kit


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Background

Versican is a large chondroitin sulfate proteoglycan with an apparent molecular mass of more than 1000kDa. In 1989, Zimmermann and Ruoslahti cloned and sequenced the core protein of fibroblast chondroitin sulfate proteoglycan. They designated it versican in recognition of its versatile modular structure. Versican belongs to the lectican protein family, with Aggrecan (abundant in Cartilage), Brevican and Neurocan (nervous system proteoglycans) as other members. Versican is also known as Chondroitin sulfate proteoglycan core protein 2 (CSPG-2), PG-M, and Chondroitin sulfate proteoglycan 2.
The role of versican in cell adhesion, migration, and proliferation is extensively studied. Versican is often considered an anti-adhesive molecule. Considering the large size (>1000 kDa) and hydration capability of versican, it is possible that the interaction of integrins (large family of cell adhesion molecules) with their cell surface receptors is sterically hindered. Expression of versican is observed in various adult tissues such as blood vessels, skin,The role of versican in cell adhesion, m and developing heart. Smooth muscle cells of blood vessels, epithelial cells of skin, and the cells of central and peripheral nervous system are a few examples of cell types that express versican physiologically. Versican is involved in development, guiding embryonic cell migration important in the formation of the heart and outlining the path for neural crest cell migration. Versican is a key factor in inflammation through interactions with adhesion molecules on the surfaces of inflammatory leukocytes and interactions with chemokines that are involved in recruiting inflammatory cells. Increased versican expression is often observed in tumor growth in tissues such as breast, brain, ovary, gastrointestinal tract, prostate, and melanoma, Sarcoma, and mesothelioma. In the adult central nervous system, versican is found in perineuronal nets, where it may stabilize synaptic connections. Versican can also inhibit nervous system regeneration and axonal growth following an injury to the central nervous system.

Feature and Advantages

<PRINCIPLE OF THE ASSAY>
The microtiter plate provided in this kit has been pre-coated with an antibody specific to Versican. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated polyclonal antibody preparation specific for Versican 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 Versican, 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 Versican 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 human Versican. No significant cross-reactivity or interference was observed.

<DETECTION RANGE>
1.56 ng/ml -100 ng/ml. The standard curve concentrations used for the ELISA’s were 100ng/ml, 50 ng/ml, 25 ng/ml, 12.5 ng/ml, 6.25 ng/ml, 3.12 ng/ml, 1.56 ng/ml.

<SENSITIVITY>
The minimum detectable dose of human Versican is typically less than 0.39 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.

Composition

   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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Assay range:1.56 ng/ml -100 ng/ml.
Assay sample volume:100 µL (Cell Culture Supernates, Plasma,Serum)
Assay time:6-8 hr. {3 hr


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