iVIEW™ DAB Detection Kit from Ventana

May 08, 2013

Overall

Quality of Results

Ease-of-Optimization

What do these ratings mean?
Write a Review
Newcastle University
Academic Haematology
Research Associate

Bcl-2 IHC staining in diffuse large B-cell lymphoma cells. The antibody demonstrates cytoplasmic and membrane staining of large CD20-positive malignant cells.

Company:

Ventana

Product Name:

iVIEW™ DAB Detection Kit

Catalog Number:

760-091

Info

Image
  • Company: Ventana
  • Product Name: iVIEW™ DAB Detection Kit
  • Catalog Number: 760-091

The iVIEW™ DAB Detection Kit is designed for automated immunohistochemical (IHC) staining of paraffin embedded or frozen tissue sections. The kit specifically detects mouse IgG, IgM and rabbit IgG antibodies bound to antigen and the specific antibody is located by a biotin-conjugated secondary antibody. Following this, a streptavidin enzyme (streptavidin-HRP) conjugates with the biotin-bound secondary antibody and visualization is made possible via hydrogen peroxide substrate and DAB, producing a brown precipitate. The iVIEW DAB Detection Kit is designed for use in conjunction with the Ventana® BenchMark or NexES automated systems and contains 6 pre-diluted and ready-to-use dispensers comprising Inhibitor, Biotinylated Ig Secondary Antibody, Streptavidin-HRP, Hydrogen Peroxide, DAB Substrate and Copper (250 tests). The primary antibody can be either commercially supplied in a pre-diluted dispenser or manually applied during a titration step.

Experimental Design and Results Summary

Application

I have used the iVIEW DAB Detection Kit to stain for the presence of various proteins in slides cut from formalin fixed paraffin embedded (FFPE) lymphoma tissue. These are then manually scored using the percentage of positively staining malignant cells by histopathologist specialists.

Starting Material

4 micron sections cut from FFPE tissue and mounted on SuperFrost Plus slides.

Protocol Overview

Sections of FFPE biopsies were cut at 4 microns and mounted onto SuperFrost Plus slides. Labels were printed for each slide using the Ventana system; each label contains a barcode comprising all of the protocol information required. The staining is then performed by the automated Benchmark system in conjunction with the iVIEW DAB Detection Kit. Briefly, sections are run through a de-paraffinization step followed by antigen retrieval, staining, several wash steps and counterstains with haematoxylin and blueing reagents. These two reagents must be purchased separately (cat#760-2021 and #760-2037, respectively).

Tips

Some background staining may be experienced due to binding of endogenous biotin. However, Ventana supplies a separate blocking kit (cat # 760-050) which can reduce this problem.

Results Summary

Clear and crisp staining of nuclear or cytoplasmic proteins.

Additional Notes

None

Image Gallery

Bcl-2 IHC staining in diffuse large B-cell lymphoma cells. The antibody demonstrates cytoplasmic and membrane staining of large CD20-positive malignant cells.

Summary

The Good

Reliable kit for automated IHC staining and less expensive than some of the alternative kits.

The Bad

Problems with background staining can be experienced with some antibodies.

The Bottom Line

Reliable kit for high quality IHC staining and good value for money.

Join the discussion