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Immunofluorescence analysis of Ubiquitin was performed using 70% confluent log phase SH-SY5Y 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 Ubiquitin Rabbit Polyclonal Antibody (PA5-16829) at 2ug/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 nuclear and cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Recombinant full-length human ubiquitin protein|
|Storage buffer||PBS, pH 7.6, with 0.2% BSA|
|Contains||15mM sodium azide|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Immunohistochemistry (Paraffin) (IHC (P))||1:50-1:200|
* 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.
PA5-16829 targets Ubiquitin in IHC (P) applications and shows reactivity with Human samples.
The PA5-16829 immunogen is recombinant full-length human ubiquitin protein.
Ubiquitin (Ub) plays a very important role in regulated non-lysosomal ATP dependent protein degradation. The protein to be degraded is conjugated to Ub and the ubiquinated protein is then selectively degraded by a 26S complex, multicatalytic cytosolic and nuclear protease termed proteasome. The Ub-proteasome proteolytic pathway, which is a complex process, is implicated to be of great importance for regulating numerous cellular processes.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
40S ribosomal protein S27a; epididymis luminal protein 112; ubiquitin and ribosomal protein S27a; ubiquitin C; ubiquitin carboxyl extension protein 80; ubiquitin-CEP80
CEP80; HEL112; RPS27A; S27A; UBA80; UBC; UBCEP1; UBCEP80