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FAQS

IMMUNOHISTOCHEMISTRY/ IMMUNOCYTOCHEMISTRY

General

Buffer Recipes

A printable pdf for the most commonly used buffers

Dilutions vs Concentrations

A quick reference chart for the most common dilutions required for Vector products.

Recommended Dilutions and Buffers

Suggested buffers for our reagents for different applications

Dehydration and Clearing after Immunostaining

A printable recommended protocol for use with Vector® Substrates

Blocking Endogenous HRP

A selection of methods for blocking peroxidase

High Temperature Antigen Unmasking

Several methods of unmasking antigens are possible, here are methods using a pressure cooker, microwave, and steamer

Trypsin Unmasking

Some epitopes require trypsin digestion to achieve the best staining.

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.  

ABC Methods

VECTASTAIN ABC Kit Protocols

All our VECTASTAIN ABC protocols in one place.

Creating a Custom ABC Kit

Our mix and match reagents are very flexible. A whole range of biotinylated reagents can be detected.

Choosing a Substrate

Please see our 'Substrate' FAQ page for further information, or the link below to our substrate properties brochure.

Trouble Shooting Guide

A guide considering sources of issues using the ABC method, and suggestions to fix them.

Polymer Detection Systems

ImmPRESS™ Polymer Protocols

Quick links to our whole range of ImmPRESS™ Polymer Detection Kits

Polymer Detection Trouble Shooting Guide

A quick guide to help solve staining problems when using a polymer detection system

Working on Rodent Tissues

Guidelines for Working on Rodent Tissue.

Mouse on Mouse, or Rat antibody on Mouse tissue & vice versa?

Help is here.

Trouble Shooting MOM

Specialist help for trouble shooting the Mouse on Mouse Kits

Multiple Labelling/ Multiplexing

Multiple Labelling

Multiple labelling can be challenging. We have put together a brochure to cover this in depth.

Multiple Labelling With ImmPRESS

Further considerations to the Multiple Labelling Guide when using the ImmPRESS reagents for multiple labelling.

Multiple Labelling with ImmPRESS Excel

Further considerations to the Multiple Labelling Guide when using the ImmPRESS Excel reagents for multiple labelling.

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.

IMMUNOFLUORESCENCE

Buffer Recipes

A printable pdf for the most commonly used buffers

Dilutions vs Concentrations

A quick reference chart for the most common dilutions required for Vector products.

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.

Fluorescent Enzyme Substrates

Some precipitating substrates can also be visible under the fluorescent microscope.

Double Immunofluorescence with Biotinylated Secondaries

An outline method for double immunofluorescence

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.

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.

Trouble Shooting MOM

Specialist help for trouble shooting the Mouse on Mouse Kits

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.  

IN SITU HYBRIDIZATION

Comparison of Anti-DIG Antibodies

 

Why DIG (Digoxigenin/Digoxin), and which one?

NEUROBIOLOGY

Suggested Protocol for Peroxidase Localization of Wheat Germ Agglutinin in Neural Tissue

Blot and Gel Detection

ABC & Western Blotting

Immunodetection of antigens on blots using the VECTASTAIN® ABC Kits - Page 2

VECTASTAIN AMP Kits Trouble Shooting Guide

Labelling 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 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.

BIOTIN & AVIDIN/ STREPTAVIDIN REAGENTS

Biotin Quantification

Our White Paper on The QuantTag Kit - a comparison to HABA & mass spectrophotometry

ENZYME SUBSTRATES

Enzyme Substrate Properties

A handy reference chart for some of the most frequently sought 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.

Substrates Spotlight

A printable on substrates - covering sensitivity, properties and combinations.

Substrates & Counterstains

Not all substrates are compatible with all counterstains, either through colour incompatibility, mounting requirements or other incompatibilities.

Substrates for Contrast with Tissue Pigments

Some tissues are pigmented, so a contrasting substrate is required.

LECTINS 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.

Suggested Protocol for Peroxidase Localization of Wheat Germ Agglutinin in Neural Tissue

Specificity of Jacalin Binding

Enrichment of NK Cells

Isolation of NK cells using Erythrina cristagalli plates

Detection of Glycoproteins

Lectin methods for Histochemistry, ELISA, and Western Blot Applications

Still have a question? E-mail or phone us!

Independent reviews of our products can be found on Biocompare.com and selectscience.net.