Chicken Immunoglobulin A,IgA ELISA Kit

Instructions
Code CSB-E11232Ch
Size 96T,5×96T,10×96T
Trial Size 24T ELISA kits trial application
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Product Details

Description

The sensitive quantitative measurement of total chicken Immunoglobulin A (IgA) antigen in serum, plasma is easily performed with this 96 well strip format ELISA kit. This assay employs the competitive inhibition enzyme immunoassay technique. The microtiter plate provided in this kit has been pre-coated with IgA. Standards or samples are added to the appropriate microtiter plate wells with HRP-conjugated antibody specific for IgA. The competitive inhibition reaction is launched between with pre-coated IgA and IgA in samples or standards for HRP-conjugates IgA antibody. After thorough washing, the TMB substrate solution is added to the wells, and the color develops in opposite to the amount of IgA in the sample. The color development is stopped upon the addition of the stop solution and the intensity of the color is measured by a microplate reader at 450 nm.

This Chicken IgA ELISA Kit has high sensitivity and excellent specificity for detection of IgA. And it also has been validated with precision low than 10%, good linearity, high recovery, and lot-to-lot consistency. Refer to the product instructions for more details.

IgA is the most abundant antibody present on mucosal surfaces where the pathogens enter the body. It plays a critical role in protecting from mucosal pathogens. The production of IgA exceeds the synthesis of all other Ig classes. IgA closely interacts with the intestinal microbiota to increase its diversity. And IgA has a passive protective role via immune exclusion. Additionally, inhibitory ITAM signaling via the IgA Fc receptor (FcαRI/CD89) by monomeric IgA may play a role in maintaining homeostasis. Conversely, IgA immune complexes effectively activate immune cells via cross-linking inhibitors, thereby inducing pro-inflammatory responses and clearing pathogens. The fact that several pathogens developed mechanisms to break down IgA or evade FcαRI-mediated activation of immune cells highlights the importance of IgA in the removal of pathogens. Enhanced or abnormal presence of IgA immune complexes can lead to overactivation of neutrophils, which can lead to severe tissue damage in a variety of inflammatory or autoimmune diseases.

Target Name Immunoglobulin A,IgA
Abbreviation IgA
Species Gallus gallus (Chicken)
Sample Types serum, plasma
Detection Range 781.25 ng/mL-50000 ng/mL
Sensitivity 390 ng/mL
Assay Time 1-5h
Sample Volume 50-100ul
Detection Wavelength 450 nm
Research Area Immunology
Assay Principle quantitative
Measurement Competitive
Precision
Intra-assay Precision (Precision within an assay): CV%<8%      
Three samples of known concentration were tested twenty times on one plate to assess.  
Inter-assay Precision (Precision between assays): CV%<10%      
Three samples of known concentration were tested in twenty assays to assess.    
             
Linearity
To assess the linearity of the assay, samples were spiked with high concentrations of chicken IgA in various matrices and diluted with the Sample Diluent to produce samples with values within the dynamic range of the assay.
  Sample Serum(n=4)  
1:100 Average % 97  
Range % 93-105  
1:200 Average % 94  
Range % 91-98  
1:400 Average % 102  
Range % 98-106  
1:800 Average % 88  
Range % 82-96  
Recovery
The recovery of chicken IgA spiked to levels throughout the range of the assay in various matrices was evaluated. Samples were diluted prior to assay as directed in the Sample Preparation section.
Sample Type Average % Recovery Range  
Serum (n=5) 96 92-102  
EDTA plasma (n=4) 102 98-108  
             
             
Typical Data
These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed.
ng/ml OD1 OD2 Average    
50000 0.072 0.071 0.072    
25000 0.134 0.136 0.135    
12500 0.231 0.242 0.237    
6250 0.426 0.435 0.431    
3125 0.666 0.675 0.671    
1562.5 0.938 0.927 0.933    
781.25 1.248 1.147 1.198    
0 1.695 1.594 1.645    
Materials provided
  • A 96-well Assay plate --The 96-well plate has been pre-coated with chicken IgA.
  • Standard(10 x concentrate) (1 x 200 μl) --Dilute the standard at dilution series, read the OD values, and then draw a standard curve.
  • HRP-conjugated IgA antibody(100 x concentrate) (1 x 60 μl) --Bind to the IgA, and HRP catalyzes the TMB to elicit a chromogenic reaction.
  • HRP-conjugate Diluent (1 x 10 ml) --Dilute the HRP-conjugated IgA antibody solution.
  • Sample Diluent (2 x 20 ml) --Reconstitute the standard and dilute the sample to an appropriate concentration.
  • Wash Buffer (25x concentrate) (1 x 20 ml) --Wash away unbound or free substances.
  • TMB Substrate (1x 10 ml) --Act as the chromogenic agent. TMB interacts with HRP, eliciting the solution turns blue.
  • Stop Solution (1 x 10ml) --Stop the color reaction. The solution color immediately turns from blue to yellow.
  • Four Adhesive Strips (For 96 wells)
  • An Instruction manual
Materials not provided
  • A microplate reader capable of measuring absorbance at 450 nm, with the correction wavelength set at 540 nm - 570 nm.
  • An incubator that can provide stable incubation conditions up to 37°C±5°C.
  • Centrifuge
  • Vortex
  • Squirt bottle, manifold dispenser, or automated microplate washer
  • Absorbent paper for blotting the microtiter plate
  • 50-300ul multi-channel micropipette
  • Pipette tips
  • Single-channel micropipette with different ranges
  • 100ml and 500ml graduated cylinders
  • Deionized or distilled water
  • Timer
  • Test tubes for dilution
Troubleshooting
and FAQs
ELISA kit FAQs
Storage Store at 2-8°C. Please refer to protocol.
Lead Time 3-5 working days

Citations

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 Q&A
Q:

For the kit CSB-E11232Ch, a customer asks if they have been tested with tissue samples, specifically small intestine.
Would you have this information available to share?

A:
Thanks for your inquiry.
We suggest you do a pretest if you want to test small intestine.Pls let me know if you have any further questions. Thank you.

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