Human NG2/MCSP/CSPG4 enzyme-linked immunoassay kit(one step)

CAT: EHY0162 Datasheet
Specification 96 Test
Sensitivity 0.004 ng/ml (50 μl);0.21 ng/ml (10 μl);
Standard Curve Range 0.69~500 ng/ml
Standard Curve Gradient 7 Points/3 Folds
Number of Incubations 2
Sample Volume 50 μl/10 μl
Type Fully Ready-to-Use
Operation Duration 60min
ng/ml O.D. Average Corrected
0.00 0.0144 0.0150 0.0147
0.69 0.0266 0.0269 0.0268 0.0121
2.06 0.0461 0.0456 0.0459 0.0312
6.17 0.1073 0.1186 0.1130 0.0983
18.52 0.2726 0.3004 0.2865 0.2718
55.56 0.8309 0.8644 0.8477 0.8330
166.67 2.0830 2.1660 2.1245 2.1098
500.00 3.8140 3.9010 3.8575 3.8428

Precision

Intra-assay Precision Inter-assay Precision
Sample Number S1 S2 S3 S1 S2 S3
22 22 22 6 6 6
Average(ng/ml) 1.7 8.6 30.9 1.6 8.9 31.6
Standard Deviation 0.1 0.4 2.0 0.1 0.6 1.8
Coefficient of Variation(%) 5.7 4.4 6.5 6.0 6.8 5.7

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 six times on one plate to assess intra-assay precision.

Spike Recovery

The spike recovery was evaluated by spiking 3 levels of human NG2/MCSP/CSPG4 into health human serum sample. The un-spiked serum was used as blank in this experiment.
The recovery ranged from 99% to 117% with an overall mean recovery of 108%.

Sample Values

Sample Matrix Sample Evaluated Range (ng/ml) Detectable (%) Mean of Detectable (ng/ml)
Serum 30 59.50-274.06 100 129.52

Serum/Plasma – Thirty samples from apparently healthy volunteers were evaluated for the presence of NG2/MCSP/CSPG4 in this assay. No medical histories were available for the donors.

Background: NG2/MCSP/CSPG4

NG2, also known as CSPG4, MCSP, and AN2, is a 400-500 kDa transmembrane chondroitin sulfate proteoglycan (CSPG) with a protein core of approximately 300 kDa. The extracellular region can be proteolytically shed from the cell surface. Mature human NG2 consists of a 2195 amino acid (aa) extracellular domain (ECD), a 21 aa transmembrane segment, and a 77 aa cytoplasmic domain. Within aa 1583-2224, human NG2/CSPG4 shares 83% aa sequence identity with mouse and rat CSPG4. NG2 binds to the extracellular matrix proteins Laminin, Tenascin, and Collagens II, V, and VI as well as to the growth factors FGF-2 and PDGF-AA. NG2 is expressed on glial cell progenitors known as O2A cells or NG2 glia. These cells are neuronally responsive and differentiate primarily into oligodendrocytes but also into astrocytes. NG2 associates with PDGF R alpha and the AMPA R subunit GluR2. It is up-regulated on microglial cells during inflammation and contributes to the induction of inflammatory mediators. Various CSPGs in the brain inhibit neurite outgrowth through interactions with Nogo Receptor/NgR1 and NgR3. This recombinant protein product corresponds to the last 5 CSPG repeats, a region which can independently inhibit neurite outgrowth. NG2 is also expressed on vascular mural cells and capillaries. It promotes vascular endothelial cell (EC) migration and angiogenesis through interactions with Galectin-3 and Integrin alpha 3 beta 1 on EC, Plasminogen, and Angiostatin. NG2 is also expressed on a variety of tumors where it contributes to tumor cell adhesion, motility, and invasion.

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