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Product Code: 03-96
Price: 425

  • All necessary reagents for 8x12 microwells
  • Sufficient for analysis of 44 samples in duplicate.
  • Includes full instruction manuals.


Documents

pdf document Product Sheet

pdf document MSDS

pdf document Kit Insert

Test principle

OurELISA kit is designed to detect protein A in IgG-containing solutions (e.g. monoclonal antibody preparations), in acid eluates from protein A columns, and in other liquid preparations. It is a sandwich ELISA based on microtitre strips coated with affinity-purified chicken anti-protein A IgG. Protein A from the sample is bound to the microwell. Bound protein A is detected using biotinylated chicken anti-protein A IgG. A streptavidin horseradish peroxidase conjugate detects the biotin conjugate. A substrate that reacts with horseradish peroxidase is added. Color development is due to conversion of the substrate by the conjugate. A positive result is indicated as a color change. The color can be read visually or by using a microplate photometer at 450 nm.

Sample Preparation Principle

Samples tested are often of acidic pH. At this low pH, the specific immunological detection of protein A is decreased rendering poor assay performance. This is overcome by neutralizing the samples and standards. Also, protein A may be non-specifically bound to IgG which falsely lowers the signal. To avoid and normalize the effect of this problem, IgG is added to standards. Standards and samples containing IgG are then treated to denature the IgG, thus maximizing the exposure of protein A epitopes.

Test Procedure, samples with IgG

1. Normalize samples and standards with respect to IgG.

2. Remove IgG by boiling.

3. Add 100 L of standards and samples, incubate 60 min.

4. Wash strips.

5. Add 100 L of Biotinylated anti-Protein A IgY, incubate 60 min.

6. Wash strips

7. Add 100 L of HRP conjugate, incubate 30 minutes.

8. Wash strips

9. Add 100 L of Substrate solution, incubate 10 min.

10. Add 100 L of Stop solution.

11. Read absorbance at 450 nm.

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