Multiplex Assay Kit for Mucin 5 Subtype B (MUC5B) ,etc. by CBA (Cytometric Bead Array)

MUC5-B; MUC9; MG1; Oligomeric Mucus/Gel-Forming; Mucin 5,Subtype B,Tracheobronchial; Cervical mucin; High molecular weight salivary mucin MG1; Sublingual gland mucin

Specificity

This assay has high sensitivity and excellent specificity for detection of Mucin 5 Subtype B (MUC5B) ,etc. by CBA (Cytometric Bead Array).
No significant cross-reactivity or interference between Mucin 5 Subtype B (MUC5B) ,etc. by CBA (Cytometric Bead Array) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Mucin 5 Subtype B (MUC5B) ,etc. by CBA (Cytometric Bead Array) and the recovery rates were calculated by comparing the measured value to the expected amount of Mucin 5 Subtype B (MUC5B) ,etc. by CBA (Cytometric Bead Array) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 92-99 95
EDTA plasma(n=5) 87-94 90
heparin plasma(n=5) 91-102 98

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Mucin 5 Subtype B (MUC5B) ,etc. by CBA (Cytometric Bead Array) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Mucin 5 Subtype B (MUC5B) ,etc. by CBA (Cytometric Bead Array) 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 Mucin 5 Subtype B (MUC5B) ,etc. by CBA (Cytometric Bead Array) 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) 93-102% 84-91% 78-97% 85-103%
EDTA plasma(n=5) 91-98% 88-103% 87-101% 84-102%
heparin plasma(n=5) 83-101% 88-101% 84-99% 99-105%

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

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 100μL standard or sample to each well, add 10μL magnetic beads, and incubate 120 minutes protect from light at 25°C on shaker;
4. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60 minutes protect from light at 25°C on shaker;
5. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent B, and incubate 30 minutes protect from light at 25°C on shaker;
6. Wash plate once on magnetic frame;
7. Add 100μL sheath fluid, swirl for 10 minutes, read on the machine.

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Magazine Citations
The Journal of Biological Chemistry The Extracellular Signal-regulated Kinase Mitogen-activated Protein Kinase/Ribosomal S6 Protein Kinase 1 Cascade Phosphorylates cAMP Response Element-binding Protein to Induce MUC5B Gene Expression via D-Prostanoid Receptor Signaling Jcb: 34199
The Journal of Biological Chemistry The ERK MAPK/RSK1 cascade phosphorylates CREB to induce MUC5B gene expression via DP1 receptor signaling Jcb: 247684
Respiratory Researh Pyocyanin-induced mucin production is associated with redox modification of FOXA2 PubMed: PMC3765780
Respirology Secreted mucins and airway bacterial colonization in non-CF bronchiectasis PubMed: 26172851
American Thoracic Society International Conference Abstracts Mucin Secretion In Asthmatics With Osmotic Airway Hyperresponsiveness doi:10.1164
Ann Am Thorac Soc Airway Mucin 2 Is Decreased in Patients with Severe Chronic Obstructive Pulmonary Disease with Bacterial Colonization doi:10.1513
Секреторно-экссудативная реакция слизистой носа на воздействие холодного воздуха у больных бронхиальной астмой 10.20538/1682-0363-2017-2-146-158
Allergy Th2 cytokines orchestrate the secretion of MUC 5 AC and MUC 5B in IL‐5 positive chronic rhinosinusitis with nasal polyps Pubmed:29802623
European Journal of Pharmacology  Salvianolic acid B improves airway hyperresponsiveness by inhibiting MUC5AC overproduction associated with Erk1/2/P38 signaling Pubmed:29382534
Japan Society of Immunology & Allergology in Otolaryngology インフルエンザ菌Phosphorylcholine の表出と中耳ムチンMUC5AC 及びMUC5B 産生への影響 10.5648:jjiao.36.7
Journal of Investigative Medicine The characteristics and clinical significance of mucin levels in bronchoalveolar lavage fluid of patients with interstitial lung disease Pubmed: 30573494
Mucosal Immunology ASC and NLRP3 maintain innate immune homeostasis in the airway through an inflammasome-independent mechanism Pubmed: 31278375
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