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Cell & Tissue Staining Kits

Cell & Tissue Staining Kits

Stem Cells More Differentiation With Less Variation

Get the most out of your primary antibodies!

Designed to optimize staining on cryostat, paraffin-embedded, and free-floating tissue sections, these kits provide all the reagents required to maximize your IHC experiments.

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Intended for the localization of antigens in a broad range of histological and cytological specimens, these kits may b used with primary IgG antibodies manufacturd by R&D System or by other vendors. Briefly, detection is based on the formation of the Avidin-Biotin Complex (ABC). Visualization is based on enzymatic conversion of chromogenic substrates 3,3’ Diaminobenzidine (DAB) or 3-amino-9-ethylcarbazole (AEC) into a colored precipitate (brown or red, respectively), by horseradish peroxidase (HRP) at the sites of antigen localization, which can then be viewed using bright-field microscopy.

The high detection sensitivity of R&D Systems Cell and Tissue Staining Kits is achieved by using premium quality biotinylated secondary antibodies and High Sensitivity Streptavidin-conjugated HRP (HSS-HRP). HSS is a chemical analog of Streptavidin that has little net positive charge at neutral or slightly alkaline pH and will interact only with Biotin attached to secondary antibodies. HSS-HRP shows little or no non-specific binding to phospholipids, nucleic acids, and carbohydrate binding proteins.

The LSAB Method of IHC Signal Amplification. When using biotinylated secondary antibodies, further amplification of the signal can be achieved by taking advantage of the strong affinity of Avidin and Streptavidin for Biotin. Avidin is a glycoprotein in egg white that combines stoichiometrically with Biotin. Streptavidin is purified from the bacterium Streptomyces avidinii, is not glycosylated, and exhibits lower non-specific binding than Avidin. Both proteins bind four Biotins per molecule. If a biotinylated secondary antibody is employed, the signal can be significantly amplified by subsequent incubation with labeled streptavidin-biotin (LSAB Method). Streptavidin may be conjugated to a detection enzyme (e.g. horseradish peroxidase or alkaline phosphatase) or a fluorochrome. Using this amplification method requires additional steps to prevent non-specific binding of Avidin and Biotin. All required reagents are included in our Cell & Tissue Staining Kits.

Key Benefits

  • Ready to use
  • High Sensitivity Streptavidin (HSS) increases detection sensitivity
  • Suitable for tissue sections & cytospin preparations
  • Pre-diluted reagents minimize hands-on time
  • Optimizes mouse, rabbit, rat, goat, or sheep IgG primary antibody staining
 

Kit Components for IHC Optimization


Blocking reagents

To minimize non-specific Peroxidase, Avidin, and Biotin signals

Secondary antibodies

Biotinylated and pre-diluted to minimize hands-on time

HSS-conjugated HRP

Chemically optimized Streptavidin displays little or no non-specific binding to phospholipids, nucleic acids, and carbohydrate binding proteins

Chromogen

Choose DAB (brown) or AEC (red)

 
 

Data Examples


GALNT10 in Mouse Testis. Optimized using the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (Catalog # CTS019). View larger image with full figure legend.

VSIG4 in Mouse Liver. Optimized using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (Catalog # CTS008). View larger image with full figure legend.

IA-2/PTPRN in Human Pancreas. Optimized using the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (Catalog # CTS002). View larger image with full figure legend.

NT-3 in Rat Brain. Optimized using the Anti-Goat HRP-AEC Cell & Tissue Staining Kit (Catalog # CTS009). View larger image with full figure legend.

 

Figure Legends

Polypeptide GalNac Transferase 10/GALNT10 in Mouse Testis. Polypeptide GalNac Transferase 10/GALNT10 was detected in perfusion fixed frozen sections of mouse testis using Sheep Anti-Human/Mouse Polypeptide GalNac Transferase 10/GALNT10 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7575) at 5 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counterstained with hematoxylin (blue). Specific staining was localized to Golgi organelles in spermatocytes. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

VSIG4 in Mouse Liver. V-set and immunoglobulin domain containing 4 (VSIG4) was detected in perfusion fixed frozen sections of mouse liver using Goat Anti-Mouse VSIG4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4674) at 1.7 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to Kuppfer cells. View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.

IA-2/PTPRN in Human Pancreas. Islet Cell Antigen 2 (IA-2)/Protein Tyrosine Phosphatase Receptor-type N (PTPRN) was detected in immersion fixed paraffin-embedded sections of human pancreas using Mouse Anti-Human IA-2/PTPRN Monoclonal Antibody (Catalog # MAB7906) at 15 µg/mL overnight at 4 °C. Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS002) and counterstained with hematoxylin (blue). Specific staining was localized to islets. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

NT-3 in Rat Brain. NT-3 was detected in perfusion fixed frozen sections of rat brain (Purkinje cells in the cerebellum) using 10 µg/mL Human NT-3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-267-NA) overnight at 4 °C. Tissue was stained with the Anti-Goat HRP-AEC Cell & Tissue Staining Kit (red; Catalog # CTS009) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.