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Alpha 1 Antitrypsin ELISA Assay Kit

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$530.00
SKU:
A1A35-K01

 Product Description

Package insert view pdf

 

 

Intended Use
The Eagle Biosciences Alpha-1 Antitrypsin ELISA AssayKit is intended for the quantitative determination of alpha-1 antitrypsin in serum, plasma and stool. The Alpha-1 Antitrypsin ELISA
AssayKit  is for research use only and not to be used in diagnostic procedures. 
 
Assay Principle
The Alpha-1 Antitrypsin ELISA assay kit determines human alpha-1-antitrypsin according to the “sandwich”-principle. Alpha-1 Antitrypsin 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 α-1-AT 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
antitrypsin.png
Assay Background
Alpha-1-Antitrypsin is a 52 kD glycoprotein, which is produced by the liver, intestinal macrophages, monocytes and mucous membrane cells of the gut. It belongs to the group of acute phase proteins and is one of the most important proteinase inhibitors. Alpha-1-antitrypsin inhibits, beside others, the proteinases trypsin and the elastase of neutrophils. A lack of α-1-AT leads to an enhanced proteolysis. Only a very small amount of alpha-1-antitrypsin is cleaved or resorbed in the gut. Therefore the measurement of α-1-AT in stool reflects the permeability of the gut during inflammatory processes.

The Eagle Biosciences Alpha-1 Antitrypsin ELISA
Assay Kit allows an easy, rapid and precise quantitative determination of alpha-1-antitrypsin in biological samples. The kit includes all reagents ready to use for preparation of the samples.

Recent Citations

Saleem, B;Okogbule-Wonodi, AC;Fasano, A;Magder, LS;Ravel, J;Kapoor, S;Viscardi, RM;, (2017). Intestinal Barrier Maturation in Very Low Birthweight Infants: Relationship to Feeding and Antibiotic Exposure. J. Pediatr, 18331-36.

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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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