ELISA Kit for Anti-Mannose Binding Lectin Antibody (Anti-MBL)

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 Anti-Mannose Binding Lectin Antibody (Anti-MBL).
No significant cross-reactivity or interference between Anti-Mannose Binding Lectin Antibody (Anti-MBL) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of Anti-Mannose Binding Lectin Antibody (Anti-MBL) and the recovery rates were calculated by comparing the measured value to the expected amount of Anti-Mannose Binding Lectin Antibody (Anti-MBL) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 95-104 101
EDTA plasma(n=5) 79-104 99
heparin plasma(n=5) 81-105 92

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Anti-Mannose Binding Lectin Antibody (Anti-MBL) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Anti-Mannose Binding Lectin Antibody (Anti-MBL) 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 Anti-Mannose Binding Lectin Antibody (Anti-MBL) 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) 83-97% 97-105% 93-101% 93-101%
EDTA plasma(n=5) 86-93% 93-103% 83-104% 95-103%
heparin plasma(n=5) 95-103% 80-103% 79-98% 80-101%

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
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 5 times;
5. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
6. Add 50µL Stop Solution. Read at 450nm immediately.

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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
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