ELISA Kit for Immunoglobulin M (IgM) Oryctolagus cuniculus (Rabbit) Sandwich ELISA

IGHM; Immunoglobulin Heavy Constant Mu; Ig mu chain C region

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  • ELISA Kit for Immunoglobulin M (IgM) Packages (Simulation)
  • ELISA Kit for Immunoglobulin M (IgM) Packages (Simulation)
  • ELISA Kit for Immunoglobulin M (IgM) Results demonstration
  • SEA543Rb.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 94-101 98
EDTA plasma(n=5) 85-92 88
heparin plasma(n=5) 87-94 91

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Immunoglobulin M (IgM) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Immunoglobulin M (IgM) 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 Immunoglobulin M (IgM) 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) 98-105% 97-104% 78-99% 78-92%
EDTA plasma(n=5) 94-105% 81-97% 82-90% 95-102%
heparin plasma(n=5) 87-101% 83-93% 81-93% 96-104%

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 Immunoglobulin M (IgM)

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 Immunoglobulin M (IgM). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Immunoglobulin M (IgM). 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 Immunoglobulin M (IgM), 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 Immunoglobulin M (IgM) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Citations

  • Effects of Clostridium butyricum on growth performance, immune function, and cecal microflora in broiler chickens challenged with Escherichia coli K88Pubmed: 24570422
  • Speci?c Humoral Immune Response Induced by Propionibacterium acnes Can Prevent Actinobacillus pleuropneumoniae Infection in MicePubmed:24429068
  • INFLUENCE OF ZINCOVIT DROP (NUTRITIONAL FOOD SUPPLEMENT) ON IMMUNE SYSTEM IN NORMAL AND CYCLOPHOSPHAMIDE INTOXICATED WISTAR RATSjournals:3790
  • Effects of fermented cottonseed meal on growth performance, serum biochemical parameters, immune functions, antioxidative abilities, and cecal microflora in broilers10.108:09540105.2017.1311308
  • Detection of streptococcus pyogenes antibodies in acute idiopathic urticariaISSN:1940-5901/IJCEM0053690
  • Effects of and on growth performance, immune function and volatile fatty acid level of caecal digesta in broilers10.1080:09540105.2018.1457013
  • Altered immunoglobulins (A and G) in Ghanaian patients with type 2 diabetesPubmed:29623201
  • Effects of polysaccharides from Yingshan Yunwu tea on meat quality, immune status and intestinal microflora in chickensPubmed: 32224178
  • Physicochemical characterization and immunomodulatory activity of sulfated Chinese yam polysaccharidePubmed: 33010421
  • Association Between Serum Selenium Level and Subclinical Mastitis in Dairy CattlePubmed: 32583225
  • IMMUNOSTIMULATORY EFFECT OF MORINGA OLEIFERA LEAVES EXTRACT ON CYCLOPHOSPHAMIDE-INDUCED IMMUNOSUPPRESSION IN MICE
  • Comparison of immunomodulatory effects of three polysaccharide fractions from Lentinula edodes water extracts
  • Effects of chitooligosaccharide supplementation on laying performance, egg quality, blood biochemistry, antioxidant capacity and immunity of laying hens during the …
  • A comparative study of autogenous, allograft and artificial bone substitutes on bone regeneration and immunotoxicity in rat femur defect model
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  • Celecoxib ameliorates liver cirrhosis via reducing inflammation and oxidative stress along spleen-liver axis in rats33577848
  • Baicalin relieves Mycoplasma pneumoniae infection?induced lung injury through regulating microRNA?221 to inhibit the TLR4/NF?¦ÊB signaling pathway34109422
  • Trace endotoxin in reclaimed water is only one of the risk sources in subchronic inhalation exposure34090073
  • The suppression of Brd4 inhibits peripheral plasma cell differentiation and exhibits therapeutic potential for systemic lupus erythematosus
  • Immunostimulatory Effect of Moringa oleifera Extracts on Cyclophosphamide-induced Immunosuppressed Mice
  • Dietary Supplementation of Inulin Ameliorates Subclinical Mastitis via Regulation of Rumen Microbial Community and Metabolites in Dairy Cows34494854
  • Sulfated modification enhances the immunomodulatory effect of Cyclocarya paliurus polysaccharide on cyclophosphamide-induced immunosuppressed mice through …34865774
  • Three Urban of China: A Cohort Study of Maternal-Infant Factors and HM Protein ComponentsPubmed:35438093

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