Protein A ELISA Kit
96 Tests
Cat.No: CSTBk301
Store at 2-8°C
Valid Period: six months
For research and manufacture use only!
Not for therapeutic or diagnostic applications!
Please read through entire procedure before beginning!

Intended Use 
Bioyeargene Protein A ELISA Kit is intended for use in quantitating unnatural protein A constructs such as Recombinant Alkali-tolerant Protein A. The kit is for Research and Manufacturing Use Only and is not intended for diagnostic use in humans or animals. This kit is designed to provide a validated acidifing sample treatment method to dissociate Protein A from IgG products and detect Protein A contamination to less than 78pg/mL in the presence of up to 3mg/mL quantities of humanized monoclonal antibodies.

Principle of the Assay
Protein A ELISA kit applies the quantitative sandwich enzyme immunoassay technique. The microtiter plate has been pre-coated with a monoclonal antibody specific for Protein A. Samples containing Protein A  are  first  diluted  in  the  Sample Diluent provided with the kits. The Denaturing Buffer is then added and  mixed to dissociate the Protein A from the product antibody. The samples are then reacted in microtiter strips coated with a monoclonal anti-Protein A capture antibody.  A second anti-Protein A  biotinylated monoclonal antibody is simultaneously reacted forming a sandwich complex of solid phase antibody-Protein A:biotin labeled antibody. After a wash step to remove any unbound reactants, Streptavidin labeled HRP is added to each well. After a short incubation period, the strips are then reacted with TMB substrate. The enzyme-substrate reaction is terminated by addition of a sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450 nm. A standard curve is plotted relating the intensity of the color (O.D.) to the concentration of standards. The Protein A concentration in each sample is interpolated from this standard curve. 

Materials & Equipment Required But Not Provided
1. Precision pipettors and disposable tips to deliver 10-1000 μl. A multi-channel   
pipette is desirable for large assays.
2. 100 mL and 1 liter graduated cylinders.
3. Distilled or deionized water.
4. Tubes to prepare sample dilutions.
5. Microplate reader capable of measuring absorbance at 450 nm.
6. Data analysis and graphing software. 

Note:
For research use only .