Multiplex Assay Kit for Mucin 5 Subtype B (MUC5B) ,etc. by FLIA (Flow Luminescence Immunoassay)

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

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

Specificity

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

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Mucin 5 Subtype B (MUC5B) ,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 Mucin 5 Subtype B (MUC5B) ,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%

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#:MUC5B) 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
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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