ELISA Kit for Secretory Immunoglobulin A (sIgA) Mus musculus (Mouse) Sandwich ELISA

S-IgA

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  • ELISA Kit for Secretory Immunoglobulin A (sIgA) Packages (Simulation)
  • ELISA Kit for Secretory Immunoglobulin A (sIgA) Packages (Simulation)
  • ELISA Kit for Secretory Immunoglobulin A (sIgA) Results demonstration
  • SEA641Mu.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) and the recovery rates were calculated by comparing the measured value to the expected amount of Secretory Immunoglobulin A (sIgA) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 83-92 88
EDTA plasma(n=5) 84-98 90
heparin plasma(n=5) 94-103 97

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Secretory Immunoglobulin A (sIgA) 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) 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) 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) 93-102% 98-105% 80-105% 85-92%
EDTA plasma(n=5) 89-98% 79-98% 93-104% 91-104%
heparin plasma(n=5) 86-101% 80-101% 93-101% 93-105%

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.

ELISA Kit for Secretory Immunoglobulin A (sIgA)

Test principle

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Secretory Immunoglobulin A (sIgA). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Secretory Immunoglobulin A (sIgA). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Secretory Immunoglobulin A (sIgA), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Secretory Immunoglobulin A (sIgA) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

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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