ELISA Kit for Occludin (OCLN)

Tight Junction Protein Occludin TM4 Minus

Specificity

This assay has high sensitivity and excellent specificity for detection of Occludin (OCLN).
No significant cross-reactivity or interference between Occludin (OCLN) and analogues was observed.

Precision

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

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.

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Magazine Citations
Brain Research HIV-1 Tat protein increases the permeability of brain endothelial cells by both inhibiting occludin expression and cleaving occludin via matrix metalloproteinase-9 ScienceDirect: S0006899311021408
Plos one Disruption of the Circadian Clock in Mice Increases Intestinal Permeability and Promotes Alcohol-Induced Hepatic Pathology and Inflammation PubMed: PMC3688973
Journal of Neurotrauma A new panel of blood biomarkers for the diagnosis of mild traumatic brain injury/concussion in adults PubMed: 25794137
United States Patent Application Methods for Diagnosis and Treatment of Concussion or Brain Injury 2016:0103140
Sci Rep.  Blood Occludin Level as a Potential Biomarker for Early Blood Brain Barrier Damage Following Ischemic Stroke articles:srep40331
Frontiers in Pharmacology Traditional Herbal Medicine-Derived Sulforaphene LFS-01 Reverses Colitis in Mice by Selectively Altering the Gut Microbiota and Promoting Intestinal … Pubmed:29375374
Thorax Fas activation alters tight junction proteins in acute lung injury Pubmed: 30385692
Antioxidants (Basel). The Effect of Dietary Quercetin on the Glutathione Redox System and Small Intestinal Functionality of Weaned Piglets Pubmed: 31426309
DISEASE MARKERS Peripheral blood occludin level as a biomarker for preoperative cerebral edema in patients with brain tumors Pubmed: 32884584
Burns Open Intestinal Permeability in Participants With Thermal Injury: A Case Series From a Prospective, Longitudinal Study (HESTIA)
PLoS One A comparison of the effects of monotherapy with rosuvastatin, atorvastatin or ezetimibe versus combination treatment with rosuvastatin-ezetimibe and … 34492054
CNS Neuroscience & Therapeutics Higher serum occludin after successful reperfusion Is associated with early neurological deterioration Pubmed:35338575
Impact de l'incorporation d'acides gras à chaine moyenne, de suppléments de levures et d'avoine nue sur la santé intestinale ainsi que sur la consommation …
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