Lipid Antibodies
There is a growing appreciation and awareness of lipids as more than just static membrane components. Lipids have emerged as complex players in cellular signaling, membrane trafficking, and are increasingly investigated as potential therapeutic targets. This swell of interest in lipid biology requires appropriate tools for visualizing and understanding their function Echelon Biosciences carries an array of lipid antibodies that have been validated in a range of applications including ICC and IHC.
Featured Products
Monoclonal antibody (IgM) to PI(4,5)P2. Validated for use in immunofluorescence staining applications and ELISA.
Monoclonal antibody to lysobisphosphatidic acid (LBPA) also termed bis(monoacylglyceryl)phosphate (BMP).
Monoclonal antibody (IgM) to PI(4,5)P2. Validated for use in immunofluorescence staining applications and ELISA.
Monoclonal antibody to sphingosine 1-phosphate, the product of sphingosine kinase. Validated for use in immunofluorescence staining applications and ELISA.
Lipid Immunostaining
Due to the small size of lipids compared to proteins and their presumed lack unique structural features, cell and molecular biologists are often surprised that generation of antibodies to lipids is possible.
Not only are anti-lipid antibodies feasible, there are well characterized disorders such as anti-phospholipid syndrome (APS) that are the result of endogenous production of antibodies to native lipids.
However, in terms of technique there are specific points of care that must be taken with immunostaining lipids that often aren’t considered with proteins. Visit our blog to learn more. ⇒
Due to the small size of lipids compared to proteins and their presumed lack unique structural features, cell and molecular biologists are often surprised that generation of antibodies to lipids is possible.
Not only are anti-lipid antibodies feasible, there are well characterized disorders such as anti-phospholipid syndrome (APS) that are the result of endogenous production of antibodies to native lipids.
However, in terms of technique there are specific points of care that must be taken with immunostaining lipids that often aren’t considered with proteins. Visit our blog to learn more. ↓
Tissue Staining - Immunohistochemistry
Immunohistochemistry (IHC) is a standard technique for visualizing cellular targets in tissue samples. While there are special considerations for handling tissue, the general process is similar to cell staining: fixation, blocking, and staining with primary and secondary antibodies.
A point of divergence with between protein and lipid antibodies is that antigen retrieval techniques from paraffin embedded samples are more difficult with lipid targets. This is primarily due to the heat-retrieval method which may bring samples above the phase transition temperature for some lipids and thus remove or strip them from their native membranes. Thus it is generally recommended to use frozen tissue samples when planning to stain for lipid antigens.
Antibody Generation
Generating lipid antibodies follows the same general procedures as for proteins in terms of immunizing animals and then purifying antibodies from serum samples.
In the case of lipids, their self assembly into liposomes can add some flexibility to the form of the antigen used for immunization. Liposomes can be directly injected for antibody generation without additional modifications so long as the lipid head group is sufficiently accessible for recognition by immune cells.
This is not always the case, and for some lipids synthetic forms must be conjugated to a carrier in order to illicit a sufficient and specific immune response. An example conjugate for PI(3,4,5)P3 is shown at right (adapted from Chen et al. 2002).
