CLIA Kit for Mucin 5 Subtype AC (MUC5AC)

TBM; MUC5; LeB; Oligomeric Mucus/Gel-Forming; Mucin 5,Subtypes A And C,Tracheobronchial/Gastric; Gastric mucin; Lewis B blood group antigen; Major airway glycoprotein

Specificity

This assay has high sensitivity and excellent specificity for detection of Mucin 5 Subtype AC (MUC5AC).
No significant cross-reactivity or interference between Mucin 5 Subtype AC (MUC5AC) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Mucin 5 Subtype AC (MUC5AC) and the recovery rates were calculated by comparing the measured value to the expected amount of Mucin 5 Subtype AC (MUC5AC) in samples.

Matrix Recovery range (%) Average(%)
EDTA plasma(n=5) 79-104 92
heparin plasma(n=5) 92-101 97

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Mucin 5 Subtype AC (MUC5AC) 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 AC (MUC5AC) 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 AC (MUC5AC) 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
EDTA plasma(n=5) 78-102% 86-101% 92-105% 79-94%
heparin plasma(n=5) 81-95% 93-101% 86-94% 82-95%

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
Substrate A 1×10mL Substrate B 1×2mL
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 100µL Substrate Solution. Incubate 10 minutes at 37°C;
8. Read RLU value immediately.

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Magazine Citations
Annals of surgical oncology Pancreatic cyst fluid and serum mucin levels predict dysplasia in intraductal papillary mucinous neoplasms of the pancreas. PubMed: 20717734
Investigative Ophthalmology & Visual Science Entrapment of Conjunctival Goblet Cells by Desiccation-Induced Cornification PubMed: 21421863
Cell & Molecular Biology Role of aquaporin 5 in antigen-induced airway inflammation and mucous hyperproduction in mice PubMed: 20550619
Mucosal Immunology Homeostatic control of conjunctival mucosal goblet cells by NKT-derived IL-13 Nature: 201082a
European Journal of Pharmacology Naringin attenuates EGF-induced MUC5AC secretion in A549 cells by suppressing the cooperative activities of MAPKs-AP-1 and IKKs-IκB-NF-κB signaling pathways ScienceDirect: S0014299912005523
PLoS One Baicalein Reduces Airway Injury in Allergen and IL-13 Induced Airway Inflammation PubMed: PMC3639905
Respiratory Researh Pyocyanin-induced mucin production is associated with redox modification of FOXA2 PubMed: PMC3765780
Surgery A novel serum marker for biliary tract cancer: Diagnostic and prognostic values of quantitative evaluation of serum mucin 5AC (MUC5AC) Pubmed: 24468034
Biochemical and Biophysical Research Communications The human cathelicidin LL-37 enhances airway mucus production in chronic obstructive pulmonary disease Pubmed:24291709
Journal of Proteomics Proteomic analysis of cancer and mesothelial cells reveals an increase in Mucin 5AC during ovarian cancer and peritoneal interaction Pubmed:24726482
Experimental Lung Research The human Cathelicidin LL-37 induces MUC5AC mucin production by airway epithelial cells via TACE-TGF-α-EGFR pathway Pubmed:24901072
Surgery.? Assessment of bile and serum mucin5AC in cholangiocarcinoma: Diagnostic performance and biologic significance Pubmed:25151557
Allergology International Leukotriene E 4 induces MUC5AC release from human airway epithelial NCI-H292 cells Pubmed:25838093
Respirology Secreted mucins and airway bacterial colonization in non-CF bronchiectasis PubMed: 26172851
S Isabela Casagrande Jeremias Teses: Disponiveis
American Thoracic Society International Conference Abstracts Mucin Secretion In Asthmatics With Osmotic Airway Hyperresponsiveness doi:10.1164
Deletion Polymorphism and Chronic Obstructive Pulmonary Disease ReCavia (Guinea pig )lation of Viral Infection-induced Airway Remodeling Cytokine Production by the TLR3-EGFR Signaling Pathway in Human Bronchial Epithelial Cells Pubmed:27075970
Int J Clin Exp Med The influence of long-term corneal contact lens wearing on the stability and quality of tear film in Qinghai files:ijcem0020163.pdf
Ann Am Thorac Soc Airway Mucin 2 Is Decreased in Patients with Severe Chronic Obstructive Pulmonary Disease with Bacterial Colonization doi:10.1513
Sci Rep Impact of Cigarette Smoking on Tear Function and Correlation between Conjunctival Goblet Cells and Tear MUC5AC Concentration in Office Workers Pubmed:27297822
Cytokine. Interleukin-33 induces mucin gene expression and goblet cell hyperplasia in human nasalepithelial cells. pubmed:27776277
J Virol.  Mutating the CX3C Motif in the G Protein Should Make a Live Respiratory Syncytial Virus Vaccine Safer and More Effective. pubmed:28275196
Respiratory Physiology & Neurobiology Peroxiredoxin 6 suppresses Muc5ac overproduction in LPS-induced airway inflammation through H2O2-EGFR-MAPK signaling pathway. pubmed:27884794
Journal of Immunology Research Beneficial effects of prebiotic Saccharomyces cerevisiae mannan on allergic asthma mouse models pubmed:28835901
BASIC INVESTIGATION Efficacy of Sodium Hyaluronate in Murine Diabetic Ocular Surface Diseases pubmed:28644234
International Immunopharmacology Clarithromycin inhibits TNF-α-induced MUC5AC mucin gene expression via the MKP-1-p38MAPK-dependent pathway pubmed:28550735
Секреторно-экссудативная реакция слизистой носа на воздействие холодного воздуха у больных бронхиальной астмой 10.20538/1682-0363-2017-2-146-158
Cellular Microbiology An adherent mucus layer attenuates the genotoxic effect of colibactin. pubmed:29156489
Respiratory Research IL-4-induced caveolin-1-containing lipid rafts aggregation contributes to MUC5AC synthesis in bronchial epithelial cells pubmed:28931396
Chemotherapy Oxytetracycline Inhibits Mucus Secretion and Inflammation in Human Airway Epithelial Cells. pubmed:28578323
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
Inflammation Plantamajoside Inhibits Lipopolysaccharide-Induced MUC5AC Expression and Inflammation through Suppressing the PI3K/Akt and NF-κB Signaling Pathways in … Pubmed:29349683
Pulmonary Pharmacology and Therapeutics In vivo and in vitro investigation of anti-inflammatory and mucus-regulatory activities of a fixed combination of thyme and primula extracts Pubmed:29698781
European Respiratory Society Profiling of healthy and asthmatic airway smooth muscle cells following IL-1β treatment: a novel role for CCL20 in chronic mucus hyper-secretion Pubmed:29946002
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