PGP 9.5

Immunohistochemical analysis of Paraffin Embedded Sections

CL7755AP (Anti-Human PGP 9.5) staining (10 µg/ml) of a human cerebral cortex formalin-fixed, paraffin-embedded tissue section; seen at 10x (left) and 20x (right) magnification. Staining on normal brain cortex is negative. High grade tumor (glioma) shows sharp nuclear and cytoplasmic staining in more than 70% of the lesion.

CL7756AP (Anti-Human PGP 9.5) staining (10 µg/ml) of a human cerebral cortex formalin-fixed, paraffin-embedded tissue section; seen at 20x (left) and 40x (right) magnification. Staining on normal brain cortex is negative. High grade tumor (glioma) shows sharp nuclear and cytoplasmic staining in more than 70% of the lesion.

CL7756AP (Anti-Human PGP 9.5) staining (2 µg/ml) of a human colon formalin-fixed, paraffin-embedded tissue section; seen at 20x (left) and 40x (right) magnification. Cytoplasmic/membrane staining of peripheral nerve/ganglion and stromal cells are observed.

CL7756AP (Anti-PGP 9.5) staining (0.1 µg/ml) of rat epidermis tissue sections isolated from rat paw using thermolysin. The tissue was frozen in M1 embedding matrix and cut on a cryostat to 12 µm. It was fixed in a 0.1 M solution of PBS containing 0.2% (w/v) paraformaldehyde and 0.75% (w/v) picric acid. Sections were imaged using confocal microscopy. Intraepithelial nerve staining of PGP 9.5 is observed (green); tissue was counterstained with DAPI (blue). Image provided by Michael Anderson from Oklahoma State University, Center for Health Sciences. PhD student.

Immunofluorescence analysis

Anti-PGP 9.5 (CL7755AP) staining using goat anti-rabbit Alexa-488 secondary antibody on Adult Mouse Hypothalamus Cell Line mHypoA-2/12 cells (CLU177) imaged under confocal microscope with the 40X oil objective; nuclei were counterstained with DAPI (blue).

CL7756AP (Anti-PGP 9.5) staining (1:50 dilution) of mouse brain (cortex) tissue, fixed with 4% paraformaldehyde and sectioned using a vibratome to 35 µm. Goat anti-rabbit IgG AlexaFluor®555 (1:500 dilution) was used as the secondary antibody and nuclei were counterstained with Hoechst. Sections were imaged using widefield fluorescence microscopy at 60x magnification. Images provided by Dr. Andrew Chojnacki from University of Calgary, Snyder Institute for Chronic Diseases.

Western Blot Analysis

CL7755AP (0.1µg/ml) staining of Human Cerebellum (A), Mouse Brain (B) and Rat Brain (C) lysates (35µg protein in RIPA buffer). Primary incubation was 1 hour. Detected by chemiluminescence

CL7756AP (0.01µg/ml) staining of Human Cerebellum (A), Mouse Brain (B) and Rat Brain (C) lysates (35µg protein in RIPA buffer). Primary incubation was 1 hour. Detected by chemiluminescence.

Description

Protein Gene Product (PGP) is an abundant cytoplasmic neuron and neuroendocrine-cell specific protein. In recent years, antibodies to this protein have become established as markers for neurons and cells of the diffuse neuroendocrine system (DNES). PGP 9.5 appears to belong to a family of ubiquitin-carboxy terminal hydrolase isoenzymes, but while it shows activity towards synthetic ubiquitin esters, the in vivo substrate(s) of PGP 9.5 remain unknown. Monoclonal and polyclonal antibodies against PGP9.5 will stain neuronal cell bodies and axons in the central and peripheral nervous systems, small nerve fibers in nerve tissue, neuroendocrine cells in the pituitary, thyroid pancreas etc., and also tumors of the DNES. Antibodies to this protein marker can be applied in neuropathology, in experimental studies of innervations of peripheral tissues and in studies of the development and pathology of the DNES. It is emphasized that antibodies to PGP are particularly suitable for detecting small nerve fibers in peripheral tissues.