Immunofluorescence

Kristy R. Howell, PhD

By Jamshed Arslan Pharm.D.

Why: To identify a specific organelle or another cellular structure and to mark individual cells, it is necessary to counterstain them in immunocytochemistry/immunofluorescence (ICC/IF) assays.

1. Multicolor immunofluorescence staining is best carried out by sequentially incubating cells with unlabeled-primary and labeled-secondary antibodies. However when options are limited, it may also be performed by simultaneous incubation of cells with directly labelled primary antibodies.  
   

In rodents, MAP1LC3 (Microtubule-associated protein 1 light chain 3) is expressed in the renal visceral epithelial cells, or podocytes. LC3 antibody analysis has shown the protein accumulates in its membrane-bound form, LC3II, following conversion from LC3I.

The vast majority of antibodies in our antibody catalog are suitable for Western blotting studies. Devised almost 30 years ago by W. Neal Burnette, it has become a standard assay wherever antibodies are used to detect proteins.

As a top worldwide antibody supplier, we at Novus Biologicals are constantly extending our antibody database to take advantage of the latest technology and product developments. Recently, we added several embryonic stem cell marker antibodies, conjugated for use in FACS (fluorescence activated cell sorting) assays.

Recently, we at Novus Biologicals added several embryonic stem cell marker products to our antibody catalog, validated for use in fluorescent activated cell sorting (FACS) assays. They included Cripto1, PODXL, SSEA, OCT4, Nanog, SOX2, TRA-1, TERT and GPR49/LGR5 antibodies.

Antibody conjugation involves tagging on a protein, compound or dye, in order to track its interaction with specific antigens. It is a routine procedure in immunoassays. We at Novus Biologicals have a vast number of products in our conjugated antibody catalog, to which we are constantly adding.

A conjugated antibody (also known as a tagged, loaded or labeled antibody) is one that has been attached to a substrate such as an enzyme, toxin or inorganic compound. Modern immunoassay techniques frequently use proteins conjugated with a fluorescent dye. Contemporary antibody sources sometimes supply a wide range of both polyclonal and monoclonal conjugated antibodies.