Immunofluorescence analysis of Protein Gene Product 9.5 was performed using 70% confluent log phase DU-145 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with PGP9.5 Rabbit Polyclonal Antibody (PA516825) at 2µg/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Rat|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Recombinant full-length human PGP95 protein|
|Storage buffer||PBS, pH 7.6, with 1% BSA|
|Contains||<0.1% sodium azide|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Immunohistochemistry (Paraffin) (IHC (P))||1:100-1:200|
|Western Blot (WB)||1-3 µg/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
PA5-16825 targets Protein Gene Product 9.5 in IHC (P) applications and shows reactivity with Human samples.
The PA5-16825 immunogen is recombinant full-length human PGP95 protein.
Protein gene product (PGP9.5) is a neuron specific protein, structurally and immunologically distinct from neuron specific enolase. The protein, which has a molecular weight of 27kDa, was first identified by high resolution two dimensional PAGE. Standard immunohistochemical techniques have demonstrated the presence of PGP9.5 in neurons and nerve fibres at all levels of the central and peripheral nervous system, in many neuroendocrine cells, in segments of the renal tubules, in spermatogonia and Leydig cells of the testis, in ova and in some cells of both the pregnant and non-pregnant corpus luteum. This antibody is of value as a marker for neurons and may be of particular use in the study of ubiquitinated cellular inclusions characteristic of several chronic human neurodegenerative diseases. This antibody is suitable for staining of formalin-fixed, paraffin-embedded tissues.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.