Immunofluorescent analysis of Proteasome 20S Y (green) in 3T3 cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes and blocked with 3% Blocker BSA (Product # 37525) in PBS for 30 minutes at room temperature. Cells were stained with or without Proteasome 20S Y rabbit polyclonal antibody (Product # PA1-978), at a concentration of 5ug/ml for 1 hour at room temperature, and then incubated with a Goat anti-Rabbit (H+L) Superclonal Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:1000 for 1 hour at room temperature (both panels, green). Nuclei (both panels, blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ToxInsight at 20X magnification.
|Tested species reactivity||Bovine, Human, Mouse|
|Published species reactivity||Mouse, Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic Peptides: C(41) R S G S A A D T Q A V/I A D A V T Y(58)|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Western Blot (WB)||1 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.
|Western Blot (WB)||See 2 publications below|
PA1-978 detects proteasome 20S Y from purified bovine and human 26S proteasome samples.
PA1-978 has been successfully used in Western blot and ICC/IF procedures. By Western blot, this antibody detects an ~25 kDa protein representing proteasome 20S Y from purified human 26S proteasome.
The PA1-978 immunizing peptides correspond to amino acid residues 41-58 from human proteasome 20S Y and amino acid residues 74-91 from frog. These sequences are completely conserved in mouse, rat, and zebrafish. PA1-978 immunizing peptide (Cat. # PEP-173) is available for use in neutralization and control procedures.
Proteolytic degradation is critical to the maintenance of appropriate levels of short-lived and regulatory proteins as important and diverse as those involved in cellular metabolism, heat shock and stress response, antigen presentation, modulation of cell surface receptors and ion channels, cell cycle regulation, transcription, and signalling factors. The ubiquitin-proteasome pathway deconstructs most proteins in the eukaryotic cell cytosol and nucleus. Other proteins are degraded via the vacuolar pathway which includes endosomes, lysosomes, and the endoplasmic reticulum. The 26S proteasome is an ATP-dependent, multisubunit (~31), barrel-shaped molecular machine with an apparent molecular weight of ~2.5 MDa. It consists of a 20S proteolytic core complex which is crowned at one or both ends by 19S regulatory subunit complexes. The 19S regulatory subunits recognize ubiquitinated proteins and play an essential role in unfolding and translocating targets into the lumen of the 20S subunit. The PA28/11S REG Activator protein complex functions as a proteolytic activator. This complex, consisting of alpha, beta, and gamma subunits, enhances the activity of the 20S proteolytic core. An enzymatic cascade is responsible for the attachment of multiple ubiquitin molecules to lysine residues of proteins targeted for degradation.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
The ubiquitin-like protein FAT10 mediates NF-kappaB activation.
PA1-978 was used in western blot to investigate the regulatory effect of ubiquitin-like protein FAT10 on NF-kappa B activation
|Gong P,Canaan A,Wang B,Leventhal J,Snyder A,Nair V,Cohen CD,Kretzler M,D'Agati V,Weissman S,Ross MJ||Journal of the American Society of Nephrology : JASN (21:316)||2010|
Reduced ubiquitin C-terminal hydrolase-1 expression levels in dementia with Lewy bodies.
PA1-978 was used in western blot to investigate the changes of UCHL-1 in samples from dementia patients with Lewy bodies
|Barrachina M,Castaño E,Dalfó E,Maes T,Buesa C,Ferrer I||Neurobiology of disease (22:265)||2006|