I need to do immunolabeling of a thick cryosection, around 60 µm thick. What should I do to ensure good labeling with my antibody?
One thing you should do is to permeabilize well, by including 0.1% to 0.2% Triton X-100 in all washes and incubations. Another recommendation is to extend the labeling times to overnight at 4oC for both primary antibody and secondary antibody. Finally, label the section free-floating, rather than adhered to a slide, to gain penetration for the antibody from both sides.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
What are the storage conditions for Triton X-100 Surfact-Amps Detergent Solution (Cat. No. 85111)?
After opening, we recommend storing the unused solution sealed in an air-tight vial containing minimal air space or from which the air has been purged with nitrogen gas. Frequent opening should be avoided because Triton X-100 is susceptible to autoxidation upon exposure to air.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
Can you provide the shelf life for Triton X-100 Surfact-Amps Detergent Solution (Cat. No. 85111)?
Triton X-100 Surfact-Amps Detergent Solution is covered under our general 1-year warranty and is guaranteed to be fully functional for 12 months from the date of shipment, if stored as recommended (20-25 degrees C). Please see section 8.1 of our Terms & Conditions of Sale (https://www.thermofisher.com/content/dam/LifeTech/Documents/PDFs/Terms-and-Conditions-of-Sale.pdf).
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
What are the main advantages of using Thermo Scientific Pierce Surfact-Amps detergents?
Thermo Scientific Pierce Surfact-Amps detergents are highly purified, precisely diluted (10%) formulations that are ideal for applications or assays that are sensitive to contaminants that are present in unpurified detergents. We test every batch to insure that our detergents contain less than 1.0 µeq/mL peroxides and carbonyls and package them under nitrogen, to prevent oxidization during storage.
Superior quality - lower measurable contaminant levels than other leading vendors
Accurate -precise 10% detergent solution in ultrapure water
Easy-to-use - solution is simple to dispense and dilute for use
Exceptionally pure - less than 1.0 µeq/mL peroxides and carbonyls
Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.
Are detergents denaturing or non-denaturing with respect to protein structure?
Ionic detergents, or those that carry a charge, are the most likely to be denaturing to proteins. Denaturing detergents can be anionic such as sodium dodecyl sulfate (SDS) or cationic such as ethyl trimethyl ammonium bromide. These detergents totally disrupt membranes and denature proteins by breaking protein-protein interactions through changes in the three-dimensional structure of the proteins. Nondenaturing detergents can be divided into nonionic detergents (i.e., Triton X-100), bile salts (i.e., cholate), and zwitterionic detergents (i.e., CHAPS).
Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.