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Secretory IgA (sIgA) ELISA Assay Kit

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$530.00
SKU:
SGA35-K01

 Product Description

Package insert view pdf
 
 



Reference Values (Normal)
Stool:    510 -  2040 µg/ml
Saliva:    102 – 478 µg/ml
Ref: M. Martin (Hrsg.). Gastroenterologische Aspekte in der Naturheilkunde  ISBN 3-930620-29-4; S.31

It is suggested that each laboratory establish its own normal ranges

Intended Use
The Eagle Biosciences Secretory IgA (sIgA) ELISA Assay Kit is intended for the quantitative determination of sIgA in stool and saliva. This Secretory IgA (sIgA) ELISA Kit is for Research Use Only and not be used for diagnostic procedures.
 
Assay Principle
The Secretory IgA (sIgA) ELISA
Assay kit determines human secretory IgA according to the "sandwich"-principle. sIgA in sample, standard and controls binds to antibodies, which are coated to the microtiter plate. After a washing step a peroxidase labeled detection antibody is added. A second washing step is followed by the addition of the substrate which is converted to a colored product by the peroxidase. The reaction is terminated by the addition of an acidic stop solution. The optical densities are read at 450 nm (against the reference wavelength 620 nm) in a microtiter plate reader. The sIgA concentration can be calculated from the standard curve.

1.    Washing step

  • Take out the needed strips of the microtiter plate and wash 1x with 250 µl diluted WASHBUF. Remove residual buffer by tapping the plate on absorbent paper after the washing step.

2.    Incubation samples

  • Pipette 100µl STD, CTRL and samples in duplicate in the microtiter plate.
  • The strips are covered and incubated by shaking for 60 min at room temperature (18-26 °C).
  • The reaction in each well starts immediately. Pipetting should be performed as quickly as possible. When processing many samples at once the samples should be pipetted to a separate microtiter plate (150 µl) and transferred simultaneously using a multichannel pipette.

3.    Washing step

  • Discard the content of the microwells and wash 5x with 250 µl diluted WASHBUF.  Remove residual buffer by tapping the plate on absorbent paper after the last washing step.

4.    Incubation conjugate

  • Pipette 100 µl CONJ in each microwell.
  • The strips are covered and incubated by shaking for 60 min at room temperature (18-26 °C).

5.    Washing step

  • Discard the content of the microwells and wash 5x with 250 µl diluted WASHBUF. Remove residual buffer by tapping the plate on absorbent paper after the last washing step.

6.    Incubation substrate

  • Pipette 100 µl SUB in each microwell.
  • Incubate for 10-15 min at room temperature (18-26 °C) in the dark.

7.    Stopping reaction

  • Pipette 50 µl STOP in each microwell, mix well.

8.    Reading

  • Read the absorbance at 450 nm. If the microtiter plate reader allows to use a reference wavelength use 620 or 690 nm as reference wavelength.
  • Reading should be done within 5 min after stopping reaction.
  • In case that the highest standard exceeds the range of the reader the reading should be done at 405 nm against 620 nm (690 nm).


Typical Standard Curve
siga.png
Assay Background
Secretory IgA is comprised of two immunoglobulin A molecules, which are joined by a J-protein and a secretory component. The secretory component is synthesized by epithelial cells of the mucous membrane of gastrointestinal, respiratory and urogenital tracts. Secretory IgA is also produced by the saliva, tear and mammary glands. The plasma cells in the subendothelial area of mucous membranes are releasing a complex of two IgA-molecules, which are joined over the J-protein. This complex is binding to a secretory component, located at the surface of the epithelial cell. After binding, the sIgA is transported across the cell and excreted by exocytosis.  The determination of secretory IgA (sIgA) allows a first overview of the functionality of the gastrointestinal associated immune system (GALT). At this the secretory power and the degree of stimulation of the plasma cells of the intestinal submucosa is determined.

The Eagle Biosciences Secretory IgA (sIgA) ELISA
Assay kit allows for an easy, rapid and precise quantitative determination of secretory IgA in biological samples. The kit includes all reagents ready to use for preparation of the samples.

Recent Citations

Morrow, JJ;Reinhard, KJ;, (2016). Assessing the Archaeoparasitological Potential of Quids As a Source Material for Immunodiagnostic Analyses. Korean J. Parasitol, 54(5), 605-616.

Shimizu, J;Kubota, T;Takada, E;Takai, K;Fujiwara, N;Arimitsu, N;Ueda, Y;Wakisaka, S;Suzuki, T;Suzuki, N;Bifidobacteria Abundance-Featured Gut Microbiota Compositional Change in Patients with Behcet's Disease, PLoS ONE, e0153746, 11:4, 2016. doi: 10.1371/journal.pone.0153746

 

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Eagle Biosciences, Inc.
20A NW Blvd., Suite 112
Nashua, NH 03063
Email: info@eaglebio.com
Toll Free: 866-411-8023
International: +617-419-2019

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