Human secretory immunoglobulin A,SIgA ELISA Kit

Instructions
Code CSB-E09235h
Size 96T,5×96T,10×96T
Trial Size 24T ELISA kits trial application
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Product Details

Description

The Human secretory immunoglobulin A (SIgA) ELISA Kit is used to quantitatively measure the levels of SIgA in human saliva, urine, or tear. This assay employs the quantitative sandwich enzyme immunoassay technique, in which SIgA in the samples or standards are sandwiched between pre-coated SIgA antibody and HRP-conjugated antibody specific for SIgA. Following a wash to remove any unbound reagent, the TMB substrate solution is added to the wells and color develops in proportion to the amount of SIgA bound in the initial step. The color development is stopped and the intensity of the color is measured at 450 nm via a microplate reader. This kit displays many advantages, including sensitivity, specificity, precision, linearity, and recovery. The product instructions are access to more information.

SIgA is the main immunoglobulin on mucosal surfaces of humans and many other mammals. It presents the immune system's first line of defense against mucosal pathogens and mediates host protection and pathogen neutralization upon reaching the luminal side. It plays an important role in the protection and homeostatic regulation of intestinal, respiratory, and urogenital mucosal epithelia separating the outside environment from the inside of the body. This process is known as immune exclusion, which allows for the segregation of the inside of the body that needs to remain sterile from the outside world rich in antigenic stimuli including those present in air, liquid, and food. SIgA is also involved in avoiding opportunistic pathogens to enter and disseminate in the systemic compartment, as well as tightly controlling the necessary symbiotic relationship existing between commensals and the host.

Target Name secretory immunoglobulin A,SIgA
Abbreviation SIgA
Species Homo sapiens (Human)
Sample Types saliva, urine, tear
Detection Range 0.012 μg/mL-50 μg/mL
Sensitivity 0.012 μg/mL
Assay Time 1-5h
Sample Volume 50-100ul
Detection Wavelength 450 nm
Research Area Others
Assay Principle quantitative
Measurement Sandwich
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.    
             
Typical Data
These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed.
μg/ml OD1 OD2 Average Corrected  
50 2.207 2.320 2.264 2.193  
12.5 2.104 2.234 2.169 2.098  
3.125 1.855 1.846 1.851 1.780  
0.781 1.539 1.551 1.545 1.474  
0.195 0.775 0.801 0.788 0.717  
0.049 0.416 0.401 0.409 0.338  
0.012 0.192 0.196 0.194 0.123  
0 0.069 0.072 0.071    
Materials provided
  • A micro ELISA plate ---The 96-well plate has been pre-coated with an anti-human SIgA antibody.
  • Two vials lyophilized standard ---Dilute a bottle of the standard at dilution series, read the OD values, and then draw a standard curve.
  • One vial HRP-conjugated SIgA antibody (100 x concentrate) (120 μl/bottle) ---Bind to the CEA in the samples or standards and react with the substrate to make the solution chromogenic.
  • Two vials Sample Diluent (20 ml/bottle)---Dilute the sample.
  • One vial Wash Buffer (25x concentrate) (20 ml/bottle) ---Wash away unbound or free substances.
  • One vial TMB substrate (10ml/bottle) ---Interact with TMB to elicit a chromogenic reaction.
  • One vial Stop Solution (10ml/bottle) ---Stop the color reaction. The solution color immediately turns from blue to yellow.
  • Four Adhesive Strips (For 96 wells) ---Cover the microplate when incubation.
  • An instruction manual
Materials not provided
  • A microplate reader capable of measuring absorbance at 450 nm, with the correction wavelength set at 540 nm or 570 nm.
  • An incubator 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

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