proteasome subunits α2, β1 β2, β5  (Enzo Biochem)

Journal: Molecular Neurobiology

Article Title: Immortalized Alzheimer’s Disease Astrocytes: Characterization of Their Proteolytic Systems

doi: 10.1007/s12035-023-03231-z

Figure Lengend Snippet: Deregulation of UPS in 3Tg-iAstro. 26S Proteasome activity in WT-iAstro (WT) and 3Tg-iAstro (AD) (panel A ) and Western blotting of ubiquitin conjugates (panel B ). Panel ( C ) 20S proteasomal activities measured in WT-iAstro (WT) and 3Tg-iAstro (AD) through enzymatic fluorimetry assays. From left to right chymotrypsin-like activity (ChT-L), trypsin-like activity (T-L), peptidyl-glutamyl peptide-hydrolyzing activity (PGPH), and branched-chain amino acid activity (BrAAP) of the 20S proteasome are reported. Activities are expressed as percentage of WT (data from three independent subgroups of both cell types at the same passage number). Panel ( D ) 20S proteasome subunits composition. Representative Western blots of 20S proteasome constitutive (β5, β2, β1) and inducible (β5i, β2i, β1i) subunits in WT-iAstro (WT) and 3Tg-iAstro (AD) immortalized murine astrocytes. Equal protein loading was verified by using an anti-β-actin antibody and normalized expression of the target protein is reported as arbitrary units (a.u.). The densitometry from five separate blots and an immunoblot which is representative of three distinct cellular subgroups are reported. Equal protein loading was verified by using an anti-β-actin antibody and normalized expression of the target proteins is reported as arbitrary units (a.u.). Data points marked with asterisks are statistically significant compared to the WT counterpart (at the same passage number) (* p < 0.05, ** p < 0.01), upon Student’s t -test

Article Snippet: Monoclonal antibodies against β5, β2, and β1 proteasomal subunits, ubiquitin, beclin-1, LAMP1, nuclear factor erythroid 2 (NF-E2)-related factor 2 transcription factor (Nrf2), histone deacetylase 6 (HDAC6), IFN-γ, and TNF-α were obtained from Santa Cruz Biotechnology, Inc. (Heidelberg, Germany).

Techniques: Activity Assay, Western Blot, Ubiquitin Proteomics, Expressing

Journal: Biochemical pharmacology

Article Title: UBXN2A regulates nicotinic receptor degradation by modulating the E3 ligase activity of CHIP

doi: 10.1016/j.bcp.2015.08.084

Figure Lengend Snippet: Antibodies and reagents used in the experiments.

Article Snippet: Anti-β2 subunit of 20S proteasome , Biomol , 1:1000.

Techniques: Mouse Assay, Generated, Recombinant, Ubiquitin Proteomics

Journal: Molecular cell

Article Title: Activity-dependent degradation of the nascentome by the neuronal membrane proteasome

doi: 10.1016/j.molcel.2018.06.013

Figure Lengend Snippet: (A) IEG products are degraded by the NMP during bicuculline stimulation in a translation-dependent and transcription-independent manner. Experimental timeline shown above. Cycloheximide (CHX) added during proteasome inhibition. Actinomycin D (ActD) added for entire hour. Neuronal lysates were immunoblotted. Representative immunoblots shown. Data are presented as mean ± SEM (n = 3).

Article Snippet: Proteasomes were subsequently immunoprecipitated using resin conjugated to an antibody against the β2 proteasome subunit (Enzo).

Techniques: Inhibition, Western Blot

Journal: Molecular cell

Article Title: Activity-dependent degradation of the nascentome by the neuronal membrane proteasome

doi: 10.1016/j.molcel.2018.06.013

Figure Lengend Snippet: Highlights :

Article Snippet: Proteasomes were subsequently immunoprecipitated using resin conjugated to an antibody against the β2 proteasome subunit (Enzo).

Techniques: Recombinant, Protease Inhibitor, Purification, Labeling, Software

Journal: Molecular cell

Article Title: Activity-dependent degradation of the nascentome by the neuronal membrane proteasome

doi: 10.1016/j.molcel.2018.06.013

Figure Lengend Snippet: (A) IEG products are degraded by the NMP during bicuculline stimulation in a translation-dependent and transcription-independent manner. Experimental timeline shown above. Cycloheximide (CHX) added during proteasome inhibition. Actinomycin D (ActD) added for entire hour. Neuronal lysates were immunoblotted. Representative immunoblots shown. Data are presented as mean ± SEM (n = 3).

Article Snippet: Mouse anti-β2 proteasome subunit (MCP168) , Enzo Life Sciences , Cat# PW8145; RRID AB_2052386.

Techniques: Inhibition, Western Blot

Journal: Molecular cell

Article Title: Activity-dependent degradation of the nascentome by the neuronal membrane proteasome

doi: 10.1016/j.molcel.2018.06.013

Figure Lengend Snippet: Highlights :

Article Snippet: Mouse anti-β2 proteasome subunit (MCP168) , Enzo Life Sciences , Cat# PW8145; RRID AB_2052386.

Techniques: Recombinant, Protease Inhibitor, Purification, Labeling, Software