ELISA Kit for Mucin 1 (MUC1)

CD227; EMA; H23AG; KL-6; PEM; PEMT; PUM; CA15-3; CA153; Cancer antigen 15-3; Carcinoma-associated mucin; Episialin; Peanut-reactive urinary mucin; Polymorphic epithelial mucin

Specificity

This assay has high sensitivity and excellent specificity for detection of Mucin 1 (MUC1).
No significant cross-reactivity or interference between Mucin 1 (MUC1) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Mucin 1 (MUC1) and the recovery rates were calculated by comparing the measured value to the expected amount of Mucin 1 (MUC1) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 95-102 98
EDTA plasma(n=5) 93-105 101
heparin plasma(n=5) 80-97 80

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Mucin 1 (MUC1) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Mucin 1 (MUC1) 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 1 (MUC1) 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) 95-105% 79-101% 84-93% 82-89%
EDTA plasma(n=5) 94-102% 93-101% 85-92% 93-101%
heparin plasma(n=5) 97-105% 78-91% 86-96% 98-105%

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
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
Annals of surgical oncology Pancreatic cyst fluid and serum mucin levels predict dysplasia in intraductal papillary mucinous neoplasms of the pancreas. PubMed: 20717734
J Hum Lact Host Defense Proteins in Breast Milk and Neonatal Yeast Colonization PubMed: 26116637
Breast Activated Leukocyte Cell Adhesion Molecule (ALCAM) in Saudi Breast Cancer Patients as Prognostic and Predictive Indicator PubMed: 26446295
PLoS One. The Presence and Anti-HIV-1 Function of Tenascin C in Breast Milk and Genital Fluids Pubmed:27182834
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
Talanta Ultrasensitive detection of mucin 1 biomarker by immuno-loop-mediated isothermal amplification. pubmed:28107977
Cellular Microbiology An adherent mucus layer attenuates the genotoxic effect of colibactin. pubmed:29156489
Bio-protocol Enhancement of Mucus Production in Eukaryotic Cells and Quantification of Adherent Mucus by ELISA 10.21769:BioProtoc.2879
Biosensors and Bioelectronics Aptasensors as a new sensing technology developed for the detection of MUC1 mucin: A review
Journal of Electroanalytical Chemistry A highly sensitive label-free electrochemical immunosensor based on AuNPs-PtNPs-MOFs for nuclear matrix protein 22 analysis in urine sample
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