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Immunohistochemistry Resource Guide

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Table of Contents

Enzyme Substrate Combinations

Recommended enzyme substrate combinations and the recommended order in which they should be used.

Substrate Combinationscrop Substrate Combinationscrop Vectorlabs Substrate combinationscrop Vectorlabs
Key: Alkaline Phosphatase Peroxidase + Indicates good contrast – Indicates incompatiblity of substrates for various reasons
Breast Carcinoma Brown Breast Carcinoma Brown Vectorlabs Breast carcinoma brown Vectorlabs
Breast Carcinoma: • Estrogen Receptor (m), VECTASTAIN® Elite® ABC Kit, Vector® NovaRED® HRP substrate (red) • CD34 (m), VECTASTAIN ABC-AP Kit, Vector Blue AP Substrate (blue) • Cytokeratin 8/18 (m), VECTASTAIN Elite ABC Kit, Vector DAB HRP Substrate (brown).
Tonsil Blue Gray Tonsil Blue Gray Vectorlabs Tonsil blue gray Vectorlabs
Tonsil: • CD3 (m), ImmPRESS® Anti-Mouse IgG Reagent, Vector VIP HRP Substrate (purple) • Ki67 (m), ImmPRESS Anti-Mouse IgG Reagent, Vector SG HRP Substrate (blue/gray).
Prostate Red Prostate Red Vectorlabs Prostate red Vectorlabs
Prostate: • Cytokeratin 5 (m), VECTASTAIN Universal ABC-AP Kit, Vector Blue AP Substrate (blue) • CD34 (m), VECTASTAIN Universal ABC-AP Kit, Vector Red AP Substrate (red).

Multiple Labeling Examples

Colon Cancer Red Colon Cancer Red Vectorlabs Colon cancer red Vectorlabs
Colon Cancer: • Ki67 (rm), ImmPRESS-AP Anti-Rabbit IgG Reagent, Vector Blue AP Substrate (blue) • Cox2 (rm), ImmPRESS-AP Anti-Rabbit IgG Reagent, ImmPACT® Vector Red AP Substrate (red).
Colon Sg Hrp Substrate Gray Colon Sg Hrp Substrate Gray Vectorlabs Colon sg hrp substrate gray Vectorlabs
Colon: • Ki67 (rm), ImmPRESS Universal (Anti-Mouse/Anti-Rabbit IgG) HRP Reagent, Vector VIP (purple) • Cytokeratin (m), ImmPRESS Universal (Anti- Mouse/Anti-Rabbit IgG) HRP Reagent, Vector SG HRP Substrate (gray).
Breast Carcinoma Vector Vip Purple Breast Carcinoma Vector Vip Purple Vectorlabs Breast carcinoma vector vip purple Vectorlabs
Breast Carcinoma: • Ki67 (rm), ImmPRESS Reagent (HRP; Universal), Vector DAB (brown) • CD34 (m), ImmPRESS Reagent (HRP; Universal), Vector VIP (purple).
Melanoma Brown Pigments Melanoma Brown Pigments Vectorlabs Melanoma brown pigments Vectorlabs
Melanoma: • Cyclin A (m), ImmPRESS-AP Anti-Mouse IgG, ImmPACT Vector Red AP Substrate (magenta) • Melanoma Marker (m) ImmPRESS HRP Anti- Mouse IgG, Vector SG HRP Substrate (gray). Note contrast of double stain with the brown pigments in the tissue.
Tonsil Vector Vip Purple Tonsil Vector Vip Purple Vectorlabs Tonsil vector vip purple Vectorlabs
Tonsil: • CD3 (m), VECTASTAIN Elite ABC Kit (Universal), Vector DAB (brown) • CD20 (m), VECTASTAIN Elite ABC Kit (Universal), Vector SG (blue-gray) • Multi-Cytokeratin (m), VECTASTAIN Elite ABC Kit (Universal), Vector VIP (purple).
Breast Carcinoma Hrp Substrate Gray Black Breast Carcinoma Hrp Substrate Gray Black Vectorlabs Breast carcinoma hrp substrate gray black Vectorlabs
Breast Carcinoma: • Estrogen Receptor (rm), ImmPRESS Universal Reagent, Vector NovaRED HRP Substrate (red) • M2A Antigen (m), ImmPRESS Universal HRP Reagent, Vector DAB+Ni HRP Substrate (gray/black).
Colon Vector Blue Ap Substrate Blue Colon Vector Blue Ap Substrate Blue Vectorlabs Colon vector blue ap substrate blue Vectorlabs
Colon: • M2A Antigen (m), ImmPRESS Universal (Anti-Mouse/Rabbit IgG) HRP Reagent, Vector NovaRED HRP Substrate (red) • CD34 (m), VECTASTAIN Universal ABC-AP kit, Vector Blue AP Substrate (blue).
Colon Carcinoma Sg Hrp Substrate Blue Gray Colon Carcinoma Sg Hrp Substrate Blue Gray Vectorlabs Colon carcinoma sg hrp substrate blue gray Vectorlabs
Colon Carcinoma: • S100 (rp), VECTASTAIN Universal ABC-AP Kit, Vector Red AP Substrate (red) • Cytokeratin 8/18 (m), VECTASTAIN Universal Elite ABC Kit, Vector SG HRP Substrate (blue/gray).