Antibody&Beyond Protocol Guide

 

 

A Guide to Multiple Antigen Labeling - In general, immunoenzymatic methods can be used to stain two or more antigens in the same tissue section when the antigens are located in different cell types or different compartments of the same cell. In cases where the two antigens are co-localized in the same compartment of the same cell, e.g., two nuclear antigens, immunofluorescence techniques may be preferred. Please click on the links below for information and protocols.

Immunoprecipitation - Immunoprecipitation involves the interaction between a protein and its specific antibody, the separation of these immune complexes with Protein G or Protein A, and the subsequent analysis by SDS-PAGE. This technique provides a rapid and simple means to separate a specific protein from whole cell lysates or culture supernatants. Additionally, one can use immunoprecipitation to confirm the identity or study biochemical characteristics, post-translational modifications, and expression levels of a protein of interest. The procedure can be divided into several stages.

Protocols and Troubleshooting Tips - A beginner's guide to western blotting, protocols for stripping for reprobing, phosphorylated proteins and troubleshooting tips. Detailed protocol, tips and troubleshooting for immunoprecipitation. Protocols and troubleshooting tips for both direct and indirect FACS / flow cytometry. rotocols for IHC on paraffin and frozen sections, cells in culture and double IF staining (sequentially or simultaneously). Protocols and tips for direct, indirect and sandwich ELISA. Protocols and troubleshooting tips for ELISPOT. Protocols, troubleshooting and tips for successful dot blotting. Expert protocols and tips for N-ChIP and X-ChIP. Protocols and tips compiled in collaboration with experts in the Neuroscience field!

Cytokine ELISA Protocol - Due to the amplifying potential of enzyme labels, immunoassays that use enzyme-conjugated antibodies have become increasingly popular because of their high specificity and sensitivity.

Immunohistochemistry Protocol - The protocol below is written for using either purified or biotinylated primary antibodies for immunohistochemical staining of mouse frozen tissue sections.

Light Microscopic Immunocytochemistry Protocol For Tissue Culture - Most of these procedures have been developed by Dr. David M. Sherry of the University of Houston and graciously supplied to Signature Immunologics. We at Signature Immunologics have modified them specifically to address use of our IgGs in detecting amino acids. These protocols are similar to HPI protocols, but some details heve been changed to meet the requirements of working with isolated cells in polystyrene dishes.

Recommended Protocols - A set of Protocols and Applications that are recommended for use with AbD Serotec Antibodies.