ELISA Kit for Collagen Type I Alpha 1 (COL1a1) Homo sapiens (Human) Sandwich ELISA

COL-1A1; COL1-A1; COL1A-1; OI4; Collagen Alpha-1(I)chain

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  • ELISA Kit for Collagen Type I Alpha 1 (COL1a1) Packages (Simulation)
  • ELISA Kit for Collagen Type I Alpha 1 (COL1a1) Packages (Simulation)
  • ELISA Kit for Collagen Type I Alpha 1 (COL1a1) Results demonstration
  • SEA350Hu.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

Matrices listed below were spiked with certain level of recombinant Collagen Type I Alpha 1 (COL1a1) and the recovery rates were calculated by comparing the measured value to the expected amount of Collagen Type I Alpha 1 (COL1a1) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 86-97 90
EDTA plasma(n=5) 83-94 88
heparin plasma(n=5) 97-105 102

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Collagen Type I Alpha 1 (COL1a1) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Collagen Type I Alpha 1 (COL1a1) 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 Collagen Type I Alpha 1 (COL1a1) 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) 95-105% 89-97% 96-103% 82-102%
EDTA plasma(n=5) 93-101% 91-101% 90-101% 98-105%
heparin plasma(n=5) 87-95% 96-104% 88-96% 90-99%

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 Collagen Type I Alpha 1 (COL1a1)

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 Collagen Type I Alpha 1 (COL1a1). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Collagen Type I Alpha 1 (COL1a1). 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 Collagen Type I Alpha 1 (COL1a1), 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 Collagen Type I Alpha 1 (COL1a1) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Citations

  • Simvastatin-loaded porous implant surfaces stimulate preosteoblasts differentiation: an in vitro studyScienceDirect: S1079210410004919
  • Osteoblast response to puerarin-loaded porous titanium surfaces: An in vitro studyWiley: source
  • Age effects on extracellular matrix production of vocal fold scar fibroblasts in ratsPubmed: 24077847
  • Vocal Fold Fibroblast Response to Growth Factor Treatment is Age Dependent: Results From an In Vitro Study Pubmed: 24495429
  • Porphyromonas gingivalis LPS inhibits osteoblastic differentiation and promotes pro-inflammatory cytokine production in human periodontal ligament stem cells.Pubmed: 24370188
  • Establishing principles of macromolecular crowding for in vitro fibrosis research of the vocal fold lamina propriaPubmed:25545625
  • Allogeneic human dermal fibroblasts are viable in peripheral blood mononuclear co-cultureUnivmed:Source
  • The vitamin D receptor agonist, calcipotriol, modulates fibrogenic pathways mitigating liver fibrosis in-vivo: An experimental study.pubmed:27477355
  • Role of Gut-Derived Endotoxin on Type I Collagen Production in the Rat Pancreas after Chronic Alcohol Exposure pubmed:29121396
  • Prevention of Diabetic Nephropathy by Modified Acidic Fibroblast Growth Factorpubmed:28768285
  • Effect of TGFβ1, TGFβ3 and keratinocyte conditioned media on functional characteristics of dermal fibroblasts derived from reparative (Balb/c) and …Pubmed:29637306
  • LOXL2, a copper-dependent monoamine oxidase, activates lung fibroblasts through the TGF-β/Smad pathwayPubmed: 30320382
  • Association between Plasma HMGB-1 and Silicosis: A Case-Control StudyPubmed: 30558126
  • Salivary proteins from dysplastic leukoplakia and oral squamous cell carcinoma and their potential for early detectionPubmed: 31706945
  • Hesperidin inhibits the epithelial to mesenchymal transition induced by transforming growth factor-β1 in A549 cells through Smad signaling in the cytoplasm
  • Platinum nanoparticles supported on functionalized hydroxyapatite: Anti-oxidant properties and bone cells response
  • Inhibition of eEF2K synergizes with glutaminase inhibitors or 4EBP1 depletion to suppress growth of triple-negative breast cancer cells33911160
  • Antiosteoporotic Nanohydroxyapatite Zoledronate Scaffold Seeded with Bone Marrow Mesenchymal Stromal Cells for Bone Regeneration: A 3D In Vitro ModelPubmed:35682677

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