FAQS IHC/ ICCIFISHNEUROBLOTS & GELS LABELLINGAVIDIN/BIOTINSUBSTRATESLECTINSIHC/ ICCIMMUNOHISTOCHEMISTRY/ IMMUNOCYTOCHEMISTRY General Buffer Recipes A printable pdf for the most commonly used buffers Buffer Recipes Dilutions vs Concentrations A quick reference chart for the most common dilutions required for Vector products. Dilution vs Concentration Recommended Dilutions and Buffers Suggested buffers for our reagents for different applications Dilutions & Buffers Dehydration and Clearing after Immunostaining A printable recommended protocol for use with Vector® Substrates Dehydration & Clearing Blocking Endogenous HRP A selection of methods for blocking peroxidase HRP Blocking Watch the video! High Temperature Antigen Unmasking Several methods of unmasking antigens are possible, here are methods using a pressure cooker, microwave, and steamer Unmasking Protocols Watch the video! Trypsin Unmasking Some epitopes require trypsin digestion to achieve the best staining. Trypsin Digestion Finding Primary Antibodies Primary antibodies are an essential part of IHC. These dedicated search engines may help find the right primary antibody for your experiment. Please note these are external links and Vector accepts no responsibility for their content, nor does this constitute an endorsement. Linscott's Directory The Antibody Registry ABC Methods VECTASTAIN ABC Kit Protocols All our VECTASTAIN ABC protocols in one place. VECTASTAIN Protocols Watch the video! Creating a Custom ABC Kit Our mix and match reagents are very flexible. A whole range of biotinylated reagents can be detected. Create Custom ABC Choosing a Substrate Please see our 'Substrate' FAQ page for further information, or the link below to our substrate properties brochure. Substrate Properties Trouble Shooting Guide A guide considering sources of issues using the ABC method, and suggestions to fix them. Trouble Shooting Guide Polymer Detection Systems ImmPRESS™ Polymer Protocols Quick links to our whole range of ImmPRESS™ Polymer Detection Kits ImmPRESS Protocols Watch the video! Polymer Detection Trouble Shooting Guide A quick guide to help solve staining problems when using a polymer detection system Polymer TSG Working on Rodent Tissues Guidelines for Working on Rodent Tissue. Mouse on Mouse, or Rat antibody on Mouse tissue & vice versa? Help is here. Rodent on Rodent Adsorbed ImmPRESS Article Trouble Shooting MOM Specialist help for trouble shooting the Mouse on Mouse Kits M.O.M. TSG Multiple Labelling/ Multiplexing Multiple Labelling Multiple labelling can be challenging. We have put together a brochure to cover this in depth. MLB Brochure Multiple Labelling With ImmPRESS Further considerations to the Multiple Labelling Guide when using the ImmPRESS reagents for multiple labelling. ImmPRESS Multiple Label Multiple Labelling with ImmPRESS Excel Further considerations to the Multiple Labelling Guide when using the ImmPRESS Excel reagents for multiple labelling. ImmPRESS Excel Multiple Label Further references UK Training Courses If you are looking for a course to help you get started, or tackle a new application these are some of the courses we'd recommend. UK Courses References VECTASTAIN References Double Labelling References IFIMMUNOFLUORESCENCE Buffer Recipes A printable pdf for the most commonly used buffers Buffer Recipes Dilutions vs Concentrations A quick reference chart for the most common dilutions required for Vector products. Dilution vs Concentration Quenching Autofluorescence Formalin fixation can lead to autofluorescence in tissues. Although this is less likely to cause an issue if you have the luxury of a confocal microscope here are some methods that can quench this autofluorescence. Quenching Auto Fluorescence Fluorescent Enzyme Substrates Some precipitating substrates can also be visible under the fluorescent microscope. Fluorescent Substrates VECTASHIELD Mounting Media Compatibility charts and protocols VECTASHIELD Compatibility VECTASHIELD HardSet Compatibility Double Immunofluorescence with Biotinylated Secondaries An outline method for double immunofluorescence Double IF Method Double Imunofluorescence (Murine) Unlike colorimetric methods that can rely on the deposition of an enzyme substrate to provide steric hindrance to prevent interaction of subsequent layers, immunofluorescence needs careful attention to potential cross-reactivities between layers. Where 2 antibodies of murine origin are involved, or on mouse tissue we recommend this method using our M.O.M. technology. M.O.M. Double IF Amplification of a Fluorescent Signal There are several ways of achieving brighter immunofluorescence, from sensitive kits to using a biotinylated anti-(strept)avidin in a sandwich method. Sandwich Method Trouble Shooting MOM Specialist help for trouble shooting the Mouse on Mouse Kits M.O.M. TSG Finding Primary Antibodies Primary antibodies are an essential part of IHC. These dedicated search engines may help find the right primary antibody for your experiment. Please note these are external links and Vector accepts no responsibility for their content, nor does this constitute an endorsement. Linscott's Directory The Antibody Registry ISHIN SITU HYBRIDIZATION Comparison of Anti-DIG Antibodies Why DIG (Digoxigenin/Digoxin), and which one? Anti-DIG for ISH NEURONEUROBIOLOGY Suggested Protocol for Peroxidase Localization of Wheat Germ Agglutinin in Neural Tissue HRP WGA - Neuro BLOTS & GELS Blot and Gel Detection ABC & Western Blotting Immunodetection of antigens on blots using the VECTASTAIN® ABC Kits - Page 2 ABC for Blots VECTASTAIN AMP Kits Trouble Shooting Guide AMP TSG LABELLINGLabelling Reagents Labelling Nucleic Acids This 36 page guide offers guidance labelling Nucleic Acids, which method is best both for nucleotide length and future applications. Labelling Nucleic Acids Labelling Proteins and Antibodies Not all antibodies are provided commercially with a label suitable for a particular experiment. The ProtOn™ Kits are designed to add biotin or FITC simply to surface primary amino groups on proteins. This system is recommended over Thiol group labelling to retain protein binding or enzymatic activity. ProtOn Biotin ProtOn Fluorescein AVIDIN/BIOTINBIOTIN & AVIDIN/ STREPTAVIDIN REAGENTS Biotin Quantification Our White Paper on The QuantTag Kit - a comparison to HABA & mass spectrophotometry QuantTag Paper SUBSTRATESENZYME SUBSTRATES Enzyme Substrate Properties A handy reference chart for some of the most frequently sought substrate properties Substrate Properties Substrate Combinations for Multiple Labelling When multiple labelling, or multiplexing, substrate combinations can play an important role, as not all substrates are compatible together. Multiple labelling combos Substrates Spotlight A printable on substrates - covering sensitivity, properties and combinations. Substrate Spotlight Substrates & Counterstains Not all substrates are compatible with all counterstains, either through colour incompatibility, mounting requirements or other incompatibilities. Substrates & Counterstains Substrates for Contrast with Tissue Pigments Some tissues are pigmented, so a contrasting substrate is required. Pigment contrasting substrates LECTINSLECTINS AND GLYCOBIOLOGY Lectin Properties Lectins are phenomenal in their complexities of binding different carbohydrates. These pages are from a double page spread in our last catalogue. Lectin Properties Chart Suggested Protocol for Peroxidase Localization of Wheat Germ Agglutinin in Neural Tissue HRP WGA - Neuro Specificity of Jacalin Binding Jacalin binding Enrichment of NK Cells Isolation of NK cells using Erythrina cristagalli plates NK Enrichment Detection of Glycoproteins Lectin methods for Histochemistry, ELISA, and Western Blot Applications Lectin methods Vector Samples Still have a question? E-mail or phone us! Independent reviews of our products can be found on Biocompare.com and selectscience.net.