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Immunofluorescence analysis of PSMC3 was performed using 70% confluent log phase Hep G2 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 PSMC3 Rabbit Polyclonal Antibody (PA1968) 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 cytoplasmic localization. Panel e shows the control without primary antibody. The images were captured at 60X magnification.
|Tested species reactivity||Human|
|Published species reactivity||Mouse, Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Human recombinant proteasome 19S subunit S6'.|
|Purification||Ammonium sulfate precipitation|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Immunoprecipitation (IP)||Assay dependent|
|Western Blot (WB)||1:1,000|
* 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.
PA1-968 detects proteasome 19S subunit S6' from human cells.
PA1-968 has been successfully used in Western blot and immunoprecipitation procedures. By Western blot, this antibody detects a 48 kDa protein representing proteasome 19S subunit S6' from HeLa cell lysate.
PA1-968 antigen is human recombinant proteasome 19S subunit S6'.
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. Others 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. An enzymatic cascade is responsible for the attachment of multiple ubiquitin molecules to lysine residues of proteins targeted for degradation. Several genetic diseases are associated with defects in the ubiquitin-proteasome pathway. Some examples of affected proteins include those linked to cystic fibrosis, Angelman and quote;s syndrome, and Liddle syndrome.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
|Mouse||Not Cited||Comparative proteomics profiling of a phospholamban mutant mouse model of dilated cardiomyopathy reveals progressive intracellular stress responses.||Gramolini AO,Kislinger T,Alikhani-Koopaei R,Fong V,Thompson NJ,Isserlin R,Sharma P,Oudit GY,Trivieri MG,Fagan A,Kannan A,Higgins DG,Huedig H,Hess G,Arab S,Seidman JG,Seidman CE,Frey B,Perry M,Backx PH,Liu PP,MacLennan DH,Emili A||Molecular and cellular proteomics : MCP (7:519)||2008|
Proteasomal inhibition by alpha-synuclein filaments and oligomers.
PA1-968 was used in immunoprecipitation to investigate the role of alpha-synuclein filaments and alpha-synuclein oligomers in proteasomal inhibition.
|Lindersson E,Beedholm R,Højrup P,Moos T,Gai W,Hendil KB,Jensen PH||The Journal of biological chemistry (279:12924)||2004|
26S protease regulatory subunit 6A; 26S proteasome AAA-ATPase subunit RPT5; human immunodeficiency virus tat transactivator binding protein-1; proteasome (prosome, macropain) 26S subunit, ATPase, 3; Proteasome 26S subunit ATPase 3; Proteasome subunit P50; Tat-binding protein 1; TBP-1; testicular secretory protein Li 42