Multiple Antigen Labeling - Vector Laboratories

More documents

Recommendations

Info

18Skeletal Muscle (frozen, unfixed) – Double label• Alpha-Sarcoglycan (m), M.O.M. Fluorescein Kit (green).• Muscle Specific Actin (m), M.O.M. Basic Kit, Texas Red ®Avidin DCS (red).Colon (frozen) – Double label• Desmin (m), M.O.M. Fluorescein Kit (green).• Peripherin (m), M.O.M. Basic Kit, Texas Red ® Avidin DCS (red).Prostate (frozen) – Double label• Muscle Specific Actin (m), M.O.M. Fluorescein Kit (green).• Multi-Cytokeratin (m), M.O.M. Basic Kit, Texas Red ® AvidinDCS (red).Breast Carcinoma (frozen) – Double label• Estrogen Receptor (m), M.O.M. Fluorescein Kit (green).• Multi-Cytokeratin (m), M.O.M. Basic Kit, VECTASTAIN ®ABC-AP Standard Kit, Vector ® Red AP substrate (red).Tonsil (frozen) – Double label• Ki67 (m), M.O.M. Basic Kit, Texas Red ® Avidin DCS (red).• CD20 (m), M.O.M. Fluorescein Kit (green).Compare color contrast with adjacent photo.Tonsil (frozen) – Double label• Ki67 (m), M.O.M. Fluorescein Kit (green).• CD20 (m), M.O.M. Basic Kit, Texas Red ® Avidin DCS.Compare color contrast with adjacent photo.Tonsil (frozen) – Double label• p63 (m), M.O.M. Fluorescein Kit (green).• Multi-Cytokeratin (m), M.O.M. Basic Kit, Texas Red ® AvidinDCS (red).Compare color contrast with adjacent photo.Tonsil (frozen) – Double label• p63 (m), M.O.M. Basic Kit, Texas Red ® Avidin DCS (red).• Multi-Cytokeratin (m), M.O.M. Fluorescein Kit (green).Compare color contrast with adjacent photo.www.vectorlabs.com
19Appendix 1: Counterstain/Substrate Compatibility TableThis table is designed as a reference to determine the optimal counterstain/substrate combination foryour application.SubstrateCatalog No.Vector ®Hematoxylin & Vector ® Vector ®Hematoxylin QS Methyl Green Nuclear Fast RedH-3401, H-3404 H-3402 H-3403DAB (brown) SK-4100 Excellent Contrast Excellent Contrast Fair ContrastDAB-Ni (gray/black) SK-4100 Excellent Contrast Fair Contrast* Good ContrastAEC (red) SK-4200 Excellent Contrast Counterstain Incompatibility** Color IncompatibilityTMB (blue) SK-4400 Color Incompatibility Counterstain Incompatibility Excellent ContrastVECTOR ® VIP (purple) SK-4600 Fair Contrast Excellent Contrast Poor ContrastVECTOR ® SG (blue/gray) SK-4700 Poor Contrast Good Contrast Excellent ContrastVECTOR ® NovaRED (red) SK-4800 Excellent Contrast Excellent Contrast*** Color IncompatibilityVECTOR ® RED (red) SK-5100 Excellent Contrast Excellent Contrast Color IncompatibilityVECTOR ® BLACK (brown/black) SK-5200 Excellent Contrast Excellent Contrast* Excellent ContrastVECTOR ® BLUE (blue) SK-5300 Color Incompatibility Good Contrast Excellent ContrastBCIP/NBT (blue/violet) SK-5400 Color Incompatibility Excellent Contrast* Excellent ContrastGLUCOSE OXIDASE NBT (purple/blue) SK-3100 Color Incompatibility Excellent Contrast Excellent ContrastGLUCOSE OXIDASE TNBT (black) SK-3200 Excellent Contrast Excellent Contrast Excellent ContrastGLUCOSE OXIDASE INT (red/purple) SK-3300 Good Contrast Counterstain Incompatibility** Fair Contrast* This substrate shows a slight decrease in sensitivity following the Methyl Green protocol. This decrease can be minimized by reducing the heatincubation and acetone rinse times in the Methyl Green protocol.** Substrate dissolves in acetone wash.***A slight color change in Vector ® NovaRED reaction product may be seen using Methyl Green.Counterstains should be optimized for each tissue type, antigen unmasking protocol, and immunocytochemical staining intensity desired.www.vectorlabs.com
Page 1 and 2: DISCOVERYthrough color.A Guide to M
Page 3 and 4: 1Multiple Antigen Labeling in the S
Page 5 and 6: 3Peroxidase SubstratesIn general, p
Page 7 and 8: 5Order of Substrates in Labeling Pr
Page 9 and 10: 7Fig. 5Fig. 6Fig.7Add Second Primar
Page 11 and 12: 9General NotesControl SectionsContr
Page 13 and 14: 11Tonsil - Double label• CD3 (m),
Page 16 and 17: 14Immunofluorescence Staining Metho
Page 18 and 19: 16Multiple Immunofluorescent Labeli
Page 22 and 23: 20Appendix 2: Quenching Endogenous

Multiple Antigen Labeling - Vector Laboratories

Create successful ePaper yourself

Delete template?

Save as template?