Multiplex Assay Kit for Secretory Immunoglobulin A (sIgA) ,etc. by FLIA (Flow Luminescence Immunoassay) Oryctolagus cuniculus (Rabbit) Multiplex ELISA

S-IgA

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

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  • Multiplex Assay Kit for Secretory Immunoglobulin A (sIgA) ,etc. by FLIA (Flow Luminescence Immunoassay) Packages (Simulation)
  • Multiplex Assay Kit for Secretory Immunoglobulin A (sIgA) ,etc. by FLIA (Flow Luminescence Immunoassay) Packages (Simulation)
  • Multiplex Assay Kit for Secretory Immunoglobulin A (sIgA) ,etc. by FLIA (Flow Luminescence Immunoassay) Results demonstration
  • LMA641Rb.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

Matrices listed below were spiked with certain level of recombinant Secretory Immunoglobulin A (sIgA) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Secretory Immunoglobulin A (sIgA) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 80-101 93
EDTA plasma(n=5) 97-104 101
heparin plasma(n=5) 85-94 89
sodium citrate plasma(n=5) 80-92 89

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Secretory Immunoglobulin A (sIgA) ,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 Secretory Immunoglobulin A (sIgA) ,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%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Secretory Immunoglobulin A (sIgA) ,etc. by FLIA (Flow Luminescence Immunoassay) 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) 88-95% 80-90% 79-96% 84-99%
EDTA plasma(n=5) 80-102% 78-97% 83-101% 92-99%
heparin plasma(n=5) 97-105% 82-94% 85-102% 89-98%
sodium citrate plasma(n=5) 82-101% 80-99% 96-105% 89-96%

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#:sIgA) 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 50μL standard or sample to each well,
    add 10μL magnetic beads,and 50μL Detection Reagent A,incubate 60min at 37°C on shaker;
3. Wash plate on magnetic frame for three times;
4. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
5. Wash plate on magnetic frame for three times;
6. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.

Multiplex Assay Kit for Secretory Immunoglobulin A (sIgA) ,etc. by FLIA (Flow Luminescence Immunoassay)

Test principle

Analyte-specific antibodies are pre-coated onto color-coded microparticles. Microparticles, standards,Labeled antigen and samples are pipetted into wells and the immobilized antibodies bind the analytes of interest.A competitive inhibition reaction is launched between biotin labeled analytes of interest and unlabeled analytes of interest (Standards or samples) with the pre-coated antibody specific to analytes of interest. Following a wash to remove any unbound substances, Streptavidin-Phycoerythrin conjugate (Streptavidin-PE) is added to each well. A final wash removes unbound Streptavidin-PE and the microparticles are resuspended in buffer and read using the Luminex or Bio-Plex analyzer. The MFI developed is reverse proportional to the concentration of analytes of interest in the sample.

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Citations

  • Evaluation of the immune benefits of two probiotic strains Bifidobacterium animalis ssp. lactis, BB-12® and Lactobacillus paracasei ssp. paracasei, L. casei 431® in an influenza vaccination model: a randomised, double-blind, placebo-controlled study.Solgar: source
  • RESPON IMUN MUKOSA DAN SELULER PADA TIKUS YANG DIBERI BUBUK SUSU KAMBING DENGAN INFEKSI Salmonella Typhimurium [Mucosal and Cellular Immune Response of Rat Given Goat Milk Powder and Infected with Salmonella Typhimurium]Ipb: Source
  • Effects of Clostridium butyricum on growth performance, immune function, and cecal microflora in broiler chickens challenged with Escherichia coli K88Pubmed: 24570422
  • Potency of Lactobacillus plantarum Dad-13 and Sweet Potato (Ipomoea batatas) Fiber as Immunomodulator in Rats Infected With Salmonella TyphimuriumCcsenet:Source
  • Repeated Oronasal Exposure to Lipopolysaccharide Induced Mucosal IgA Responses in Periparturient Dairy CowsPubmed:25061754
  • Dietary Apostichopus japonicus enhances the respiratory and intestinal mucosal immunity in immunosuppressive micePubmed:25186733
  • Dietary squid ink polysaccharide could enhance SIgA secretion in chemotherapeutic micePubmed:25308407
  • Fecal microbiota transplantation and bacterial consortium transplantation have comparable effects on the re-establishment of mucosal barrier function in mice with intestinal dysbiosisPubMed: 26217323
  • Role of Maternal Allergy on Immune Markers in Colostrum and Secretory Immunoglobulin A in Stools of Breastfed InfantsPubMed: 26243755
  • Oropharyngeal Colostrum Administration in Extremely Premature Infants: An RCTPubMed: 25624376
  • Salivary Secretory Immunoglobulin (SIgA) and Lysozyme in Malignant Tumor Patientsjournals:8701423
  • 益生菌抵御大鼠急性腹泻的效果及其对肠道黏膜屏障的影响article:12422
  • Long-term use of ceftriaxone sodium induced changes in gut microbiota and immune system.pubmed:28220870
  • Imbalance of intestinal immune function in piglets infected by porcine circovirus type 2 during the fetal period.pubmed:28244338
  • Immunomodulatory effects of Hericium erinaceus derived polysaccharides are mediated by intestinal immunologypubmed:28266682
  • Saccharomyces boulardiiadministration can inhibit the formation of gastric lymphoid follicles induced by Helicobacter suis infectionpubmed:28115360
  • Effects of dietary threonine supplementation on intestinal barrier function and gut microbiota of laying henspubmed:28938780
  • Novel dietary polysaccharide SIP promotes intestinal secretory immunoglobulin A secretion in mice under chemotherapy10.1016/j.jff.2017.08.012
  • Evaluation of Susceptibility and Innate Immune Response to Candida albicans in Mice with Sub-healthijp.2018.689.697
  • Correlation of s-IgA and IL-6 Salivary with Caries Disease and Oral Hygiene Parameters in ChildrenPubmed: 31892186
  • Fluorescence polarization-based rapid detection system for salivary biomarkers using modified DNA aptamers containing base-appended basesPubmed: 31855403
  • Artesunate protects immunosuppression mice induced by glucocorticoids via enhancing pro-inflammatory cytokines release and bacterial clearancePubmed: 33045197
  • Effects of different doses lipopolysaccharides on the mucosal barrier in mouse intestinePubmed: 32947071
  • Yacón (Smallanthus sonchifolius) prevented inflammation, oxidative stress, and intestinal alterations in an animal model of colorectal carcinogenesisPubmed: 32567144
  • Construction and Immunogenicity Comparison of Three Virus-Like Particles Carrying Different Combinations of Structural Proteins of Avian Coronavirus Infectious?¡­33670249
  • Ingestion of High ¦Â-Glucan Barley Flour Enhances the Intestinal Immune System of Diet-Induced Obese Mice by Prebiotic Effects33799564
  • Assessment of salivary stress and pain biomarkers and their relation to self-reported pain intensity during orthodontic tooth movement: a longitudinal and prospective …34170330
  • Microarray Analysis of Paramylon, Isolated from Euglena Gracilis EOD-1, and Its Effects on Lipid Metabolism in the Ileum and Liver in Diet-Induced Obese Mice34684408
  • Saliva testing as noninvasive way for monitoring exercise-dependent response in teenage elite water polo players: A cohort study34797320
  • Fermented Corn–Soybean Meal Mixed Feed Modulates Intestinal Morphology, Barrier Functions and Cecal Microbiota in Laying Hens34827791
  • Three Urban of China: A Cohort Study of Maternal-Infant Factors and HM Protein ComponentsPubmed:35438093

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