ELISA Kit for Heparan Sulfate Proteoglycan (HSPG) Homo sapiens (Human) Sandwich ELISA

SDC2; HSPG1; SYND2; Fibroglycan; Syndecan 2; Syndecan Proteoglycan 2; Heparan Sulfate Proteoglycan 1,Cell Surface-Associated; Heparan Sulphate Proteoglycan

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  • ELISA Kit for Heparan Sulfate Proteoglycan (HSPG) Packages (Simulation)
  • ELISA Kit for Heparan Sulfate Proteoglycan (HSPG) Packages (Simulation)
  • ELISA Kit for Heparan Sulfate Proteoglycan (HSPG) Results demonstration
  • SEA565Hu.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

Matrices listed below were spiked with certain level of recombinant Heparan Sulfate Proteoglycan (HSPG) and the recovery rates were calculated by comparing the measured value to the expected amount of Heparan Sulfate Proteoglycan (HSPG) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 95-105 101
EDTA plasma(n=5) 84-103 96
heparin plasma(n=5) 84-94 90

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Heparan Sulfate Proteoglycan (HSPG) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Heparan Sulfate Proteoglycan (HSPG) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Heparan Sulfate Proteoglycan (HSPG) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 79-101% 90-99% 79-101% 97-105%
EDTA plasma(n=5) 80-88% 78-95% 91-104% 79-104%
heparin plasma(n=5) 78-101% 89-98% 94-101% 82-98%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.

ELISA Kit for Heparan Sulfate Proteoglycan (HSPG)

Test principle

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Heparan Sulfate Proteoglycan (HSPG). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Heparan Sulfate Proteoglycan (HSPG). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Heparan Sulfate Proteoglycan (HSPG), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Heparan Sulfate Proteoglycan (HSPG) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Citations

  • Cytotoxic effects of antiglypican-3 against HepG3 cell lineEbscohost: Source
  • Fibrinogen, an endogenous ligand of Toll-like receptor 4, activates monocytes in pre-eclamptic patientsPubmed: 24661950
  • Suramin inhibits hepatic tissue damage in hepatocellular carcinoma through deactivation of heparanase enzymePubmed: 24530413
  • Evaluation of antiglypican-3 therapy as a promising target for amelioration of hepatic tissue damage in hepatocellular carcinomaPubmed:25449037
  • Impairment of the Endothelial Glycocalyx in Cardiogenic Shock and its Prognostic RelevancePubmed:25692257
  • Endothelial glycocalyx layer shedding following lung resectionpubmed:27643669
  • Circulating syndecans during critical illness.pubmed:28256016
  • Cytotoxic and partial hepatoprotective activity of sodium ascorbate against hepatocellular carcinoma through inhibition of sulfatase-2 andPubmed:29669302
  • Glioma targeting peptide modified apoferritin nanocagePubmed:29726297
  • Postoperative microcirculatory perfusion and endothelial glycocalyx shedding following cardiac surgery with cardiopulmonary bypassPubmed: 30687934
  • Diet Supplementation with a Bioactive Pomace Extract from Olea europaea Partially Mitigates Negative Effects on Gut Health Arising from a Short-Term Fasting Period …
  • Heparin prevents in vitro glycocalyx shedding induced by plasma from COVID-19 patients33781826
  • A novel mutation in ext2 caused hereditary multiple exostoses through reducing the synthesis of heparan sulfate34042151
  • Response of broiler chickens fed diets supplemented with a bioactive olive pomace extract from Olea europaea to an experimental coccidial vaccine?¡­33518110
  • The effect of pre-operative methylprednisolone on postoperative delirium in elderly patients undergoing gastrointestinal surgery: a randomized, double-blind, placebo …34423832

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