ELISA Kit for Claudin 3 (CLDN3) Canis familiaris; Canine (Dog) Competition ELISA

C7orf1; CPE-R2; CPETR2; HRVP1; RVP1; Clostridium Perfringens Enterotoxin Receptor 2; Ventral Prostate.1-Like Protein; CPE-Receptor 2

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  • ELISA Kit for Claudin 3 (CLDN3) Packages (Simulation)
  • ELISA Kit for Claudin 3 (CLDN3) Packages (Simulation)
  • ELISA Kit for Claudin 3 (CLDN3) Results demonstration
  • CEF293Ca.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

Matrices listed below were spiked with certain level of recombinant Claudin 3 (CLDN3) and the recovery rates were calculated by comparing the measured value to the expected amount of Claudin 3 (CLDN3) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 96-103 101
EDTA plasma(n=5) 78-92 88
heparin plasma(n=5) 90-104 101

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Claudin 3 (CLDN3) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Claudin 3 (CLDN3) 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 Claudin 3 (CLDN3) 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) 79-89% 81-94% 99-105% 80-103%
EDTA plasma(n=5) 97-105% 86-104% 90-97% 89-103%
heparin plasma(n=5) 93-102% 78-98% 80-92% 87-94%

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 50µL standard or sample to each well.
    And then add 50µL prepared Detection Reagent A immediately.
    Shake and mix. Incubate 1 hour at 37°C;
3. Aspirate and wash 3 times;
4. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
5. Aspirate and wash 5 times;
6. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
7. Add 50µL Stop Solution. Read at 450 nm immediately.

ELISA Kit for Claudin 3 (CLDN3)

Test principle

This assay employs the competitive inhibition enzyme immunoassay technique. A monoclonal antibody specific to Claudin 3 (CLDN3) has been pre-coated onto a microplate. A competitive inhibition reaction is launched between biotin labeled Claudin 3 (CLDN3) and unlabeled Claudin 3 (CLDN3) (Standards or samples) with the pre-coated antibody specific to Claudin 3 (CLDN3). After incubation the unbound conjugate is washed off. Next, avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. The amount of bound HRP conjugate is reverse proportional to the concentration of Claudin 3 (CLDN3) in the sample. After addition of the substrate solution, the intensity of color developed is reverse proportional to the concentration of Claudin 3 (CLDN3) in the sample.

Citations

  • Gastrointestinal response and endotoxemia during intense exercise in hot and cool environmentsPubmed: 23314685
  • Effects of probiotics supplementation on gastrointestinal permeability, inflammation and exercise performance in the heatSpringer: Source
  • Clinical Characteristics Associated With Postoperative Intestinal Epithelial Barrier Dysfunction in Children With Congenital Heart Disease*PubMed: 25162512
  • Reinforcement of intestinal epithelial barrier by arabinoxylans in overweight and obese subjects: A randomized controlled trial: Arabinoxylans in gut barrier.pubmed:28214040
  • Plasma claudin-3 is associated with tumor necrosis factor-alpha-induced intestinal endotoxemia in liver diseasePubmed: 31053499
  • Intestinal Permeability in Children with Celiac Disease after the Administration of Oligofructose-Enriched Inulin into a Gluten-Free Diet—Results of a Randomized …Pubmed: 32531982
  • Markers of Intestinal Damage in Individuals with and without Obesity during a 12-Week Exercise Period.
  • Dysbiosis and Intestinal Barrier Dysfunction in Pediatric Congenital Heart Disease is Exacerbated Following Cardiopulmonary Bypass
  • Short-Term Very High Carbohydrate Diet and Gut-Training Have Minor Effects on Gastrointestinal Status and Performance in Highly Trained Endurance AthletesPubmed:35565896

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