Multiplex Assay Kit for Occludin (OCLN) ,etc. by FLIA (Flow Luminescence Immunoassay)

Tight Junction Protein Occludin TM4 Minus

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

Specificity

This assay has high sensitivity and excellent specificity for detection of Occludin (OCLN) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Occludin (OCLN) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Occludin (OCLN) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Occludin (OCLN) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 95-104 101
EDTA plasma(n=5) 94-105 102
heparin plasma(n=5) 81-91 87
sodium citrate plasma(n=5) 79-103 98

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Occludin (OCLN) ,etc. by FLIA (Flow Luminescence Immunoassay) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Occludin (OCLN) ,etc. by FLIA (Flow Luminescence Immunoassay) 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 Occludin (OCLN) ,etc. by FLIA (Flow Luminescence Immunoassay) 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) 97-104% 82-97% 88-96% 98-105%
EDTA plasma(n=5) 84-101% 95-103% 93-101% 94-105%
heparin plasma(n=5) 78-101% 93-101% 79-92% 93-102%
sodium citrate plasma(n=5) 97-105% 85-99% 91-101% 83-94%

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
96-well plate 1 Plate sealer for 96 wells 4
Pre-Mixed Standard 2 Standard Diluent 1×20mL
Pre-Mixed Magnetic beads (22#:OCLN) 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. Preparation of standards, reagents and samples before the experiment;
2. Add 100μL standard or sample to each well,
    add 10μL magnetic beads, and incubate 90min at 37°C on shaker;
3. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60min at 37°C on shaker;
4. Wash plate on magnetic frame for three times;
5. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
6. Wash plate on magnetic frame for three times;
7. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.

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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 …
Catalog No. Related products for research use of Rattus norvegicus (Rat) Organism species Applications (RESEARCH USE ONLY!)
RPC228Ra01 Recombinant Occludin (OCLN) Positive Control; Immunogen; SDS-PAGE; WB.
PAC228Ra01 Polyclonal Antibody to Occludin (OCLN) WB; IHC; ICC; IP.
MAC228Ra21 Monoclonal Antibody to Occludin (OCLN) WB; IHC; ICC; IP.
SEC228Ra ELISA Kit for Occludin (OCLN) Enzyme-linked immunosorbent assay for Antigen Detection.
LMC228Ra Multiplex Assay Kit for Occludin (OCLN) ,etc. by FLIA (Flow Luminescence Immunoassay) FLIA Kit for Antigen Detection.