Multiplex Assay Kit for Mannose Binding Lectin (MBL) ,etc. by CBA (Cytometric Bead Array)

MBL2; COLEC1; HSMBPC; MBP1; MBP; Collectin-1; Mannose-binding protein C; Mannan Binding Protein; Mannose-Binding Lectin(Protein C)2,Soluble(Opsonic Defect)

Specificity

This assay has high sensitivity and excellent specificity for detection of Mannose Binding Lectin (MBL) ,etc. by CBA (Cytometric Bead Array).
No significant cross-reactivity or interference between Mannose Binding Lectin (MBL) ,etc. by CBA (Cytometric Bead Array) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Mannose Binding Lectin (MBL) ,etc. by CBA (Cytometric Bead Array) and the recovery rates were calculated by comparing the measured value to the expected amount of Mannose Binding Lectin (MBL) ,etc. by CBA (Cytometric Bead Array) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 82-95 88
EDTA plasma(n=5) 85-93 89
heparin plasma(n=5) 89-103 97

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Mannose Binding Lectin (MBL) ,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 Mannose Binding Lectin (MBL) ,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 Mannose Binding Lectin (MBL) ,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) 79-90% 83-91% 78-93% 82-93%
EDTA plasma(n=5) 99-105% 97-104% 78-91% 91-99%
heparin plasma(n=5) 99-105% 88-104% 95-105% 92-99%

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
American Journal of Nephrology Analysis of the Urine Proteome of Human Contrast-Induced Kidney Injury Using Two-Dimensional Fluorescence Differential Gel Electrophoresis/Matrix-Assisted Laser Desorption Time-of-Flight Mass Spectrometry/Liquid Chromatography Mass Spectrometry. Karger: 000255439
Current Eye Research Serum Levels and H/L Gene Polymorphism of Mannose-Binding Lectin in Primary Open Angle Glaucoma Ingenta: art00007
Journal of Clinical Immunology Complement activation contributes to the injury and outcome of kidney in human anti-glomerular basement membrane disease. PubMed: 22941511
Clinical and Experimental Immunology Urinary mannose-binding lectin is a biomarker for predicting the progression of immunoglobulin (Ig)A nephropathy PubMed: PMC3406374
Clin J Am Soc Nephrol. Alternative Complement Pathway Activation Products in Urine and Kidneys of Patients with ANCA-Associated GN Pubmed: 24115193
ACTA TROPICA Novel findings on the role of ficolins and colectins in the innate response against Leishmania braziliensis Pubmed: 32827456
PLoS One Complement activation profile of patients with primary focal segmental glomerulosclerosis Pubmed: 32569286
LIFE SCIENCES The lectin pathway of complement and the initial recognition of Leishmania infantum promastigotes 34242658
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