Multiplex Assay Kit for Hydroxyproline (Hyp) ,etc. by FLIA (Flow Luminescence Immunoassay) Pan-species (General) Multiplex ELISA

(2S,4R)-4-Hydroxyproline; L-Hydroxyproline(C5H9O3N)

(Note: Up to 8-plex in one testing reaction)

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  • Multiplex Assay Kit for Hydroxyproline (Hyp) ,etc. by FLIA (Flow Luminescence Immunoassay) Packages (Simulation)
  • Multiplex Assay Kit for Hydroxyproline (Hyp) ,etc. by FLIA (Flow Luminescence Immunoassay) Packages (Simulation)
  • Multiplex Assay Kit for Hydroxyproline (Hyp) ,etc. by FLIA (Flow Luminescence Immunoassay) Results demonstration
  • LMA621Ge.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

Matrices listed below were spiked with certain level of Hydroxyproline (Hyp) and the recovery rates were calculated by comparing the measured value to the expected amount of Hydroxyproline (Hyp) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 92-101 95
EDTA plasma(n=5) 89-97 94
heparin plasma(n=5) 84-101 91

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Hydroxyproline (Hyp) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Hydroxyproline (Hyp) 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 Hydroxyproline (Hyp) 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) 82-102% 78-99% 78-101% 98-105%
EDTA plasma(n=5) 82-101% 87-95% 88-103% 91-105%
heparin plasma(n=5) 79-93% 90-101% 79-98% 96-105%

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
96-well plate 1 Plate sealer for 96 wells 4
Pre-Mixed Standard 2 Standard Diluent 1×20mL
Pre-Mixed Magnetic beads (22#:Hyp) 1 Analysis buffer 1×20mL
Pre-Mixed Detection Reagent A 1×120μL Assay Diluent A 1×12mL
Detection Reagent B (PE-SA) 1×120μL Assay Diluent B 1×12mL
Sheath Fluid 1×10mL Wash Buffer (30 × concentrate) 1×20mL
Instruction manual 1

Assay procedure summary

1. Add 200μL analysis buffer solution to each well of the plate for pre-wetting;
2. Preparation of standards, reagents and samples before the experiment;
3. Add 50μL standard or sample to each well, add 50μL prepared Detection Reagent A, add 10μL magnetic beads, and incubate 90 minutes protect from light at 25°C on shaker;
4. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent B, and incubate 30 minutes protect from light at 25°C on shaker;
5. Wash plate once on magnetic frame;
6. Add 100μL sheath fluid, swirl for 10 minutes, read on the machine.

Multiplex Assay Kit for Hydroxyproline (Hyp) ,etc. by FLIA (Flow Luminescence Immunoassay)

Test principle

Anti-Hydroxyproline (Hyp) antibody was coated on the surface of magnetic beads. A competitive inhibition reaction is launched between biotin labeled Hydroxyproline (Hyp) and unlabeled Hydroxyproline (Hyp) (Standards or samples) with the pre-coated antibody specific to Hydroxyproline (Hyp). PE-labeled avidin was added. The amount of bound PE conjugate is reverse proportional to the concentration of Hydroxyproline (Hyp) in the sample, the intensity of MFI developed is reverse proportional to the concentration of Hydroxyproline (Hyp) in the sample.

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Citations

  • Spleen-Derived Interleukin-10 Downregulates the Severity of High-Fat Diet-Induced Non-Alcoholic Fatty Pancreas DiseasePubMed: PMC3532347
  • Prediction of fibrosis in hepatitis C patients: assessment using hydroxyproline and oxidative stress biomarkersSpringer: Source
  • Royal jelly affects collagen crosslinking in bone of ovariectomized ratsSciencedirect:S1756464614000346
  • Tanshinone IIA ameliorates bleomycin-induced pulmonary fibrosis and inhibits transforming growth factor-beta-β–dependent epithelial to mesenchymal transitionPubMed: 25911951
  • Theophylline, an old drug with multi-faceted effects: Its potential benefits in immunological liver injury in ratsPubMed: 26165752
  • Effects of 4% Icodextrin on Experimental Spinal Epidural Fibrosispdf:JTNEPUB_15079_online.pdf
  • Celecoxib attenuates depressive-like behavior associated with immunological liver injury inC57BL/6 mice through TNF-α and NF-κb dependent mechanisms.pubmed:27580523
  • Sustained-released mixture of vascular endothelial growth factor 165 and fibrin glue strengthens healing of ileal anastomoses in a rabbit model with …doi.org:10.4174
  • Effects of panax notoginseng saponin on the pathological ultrastructure and serum IL‐6 and IL‐8 in pulmonary fibrosis in rabbitsPubmed:29932250
  • Renin–angiotensin system inhibition ameliorates CCl-induced liver fibrosis in mice through the inactivation of nuclear transcription factor kappa B10.1139:cjpp-2017-0728
  • Pirfenidone suppresses bleomycin‑induced pulmonary fibrosis and periostin expression in rats10.3892:etm.2018.6378
  • Therapeutic effect of hepatocyte growth factor-overexpressing bone marrow-derived mesenchymal stem cells on CCl 4-induced hepatocirrhosis.
  • The effects of platelet‐rich‐plasma gel application to the colonic anastomosis in hyperthermic intraperitoneal chemotherapy: An experimental rat modelPubmed: 31448554
  • Pantoea agglomerans chronic exposure induces epithelial-mesenchymal transition in human lung epithelial cells and mice lungsPubmed: 32146192
  • Is platelet-rich plasma improves the anastomotic healing in hyperthermic intraperitoneal chemotherapy with oxaliplatin: an experimental rat studyPubmed: 32051817
  • Protective Effects of Contractubex® on Stricture Formation after Experimental Corrosive Esophageal Burns in RatsPubmed: 32654881
  • Effect of Oral Channa striatus in Laparotomy Wound Healing of Malnourished Rats.
  • Handgrip Strength and Vitamin D as Predictors of Liver Fibrosis and Malnutrition in Chronic Hepatitis C Patients33884041
  • Beneficial impact of cathelicidin on hypersensitivity pneumonitis treatment¡ªIn vivo studies33999928
  • Curdione and Schisandrin C Synergistically Reverse Hepatic Fibrosis via Modulating the TGF-β Pathway and Inhibiting Oxidative Stress34858986

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