Immunofluorescence analysis of PSMD2 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 PSMD2 Rabbit Polyclonal Antibody (PA1964) at 1:250 dilution 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 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytosolic localization. Panel e shows the control without primary antibody. The images were captured at 60X magnification.
|Tested species reactivity||Human|
|Published species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Human recombinant proteasome 19S subunit S2.|
|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 *|
|Immunoprecipitation (IP)||Assay dependent|
|Western Blot (WB)||1:250|
* 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.
PA1-964 detects proteasome 19S subunit S2 from human cells.
PA1-964 has been successfully used in Western blot and immunoprecipitation procedures. By Western blot, this antibody detects a 100 kDa protein representing proteasome 19S subunit S2 in HeLa cell lysate. This antibody detects, to a lesser extent an ~50-kDa protein which could correspond to subunit degradation product.
PA1-964 antigen is recombinant human proteasome 19S subunit S2.
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.
The Proteasome Ubiquitin Receptor hRpn13 and Its Interacting Deubiquitinating Enzyme Uch37 Are Required for Proper Cell Cycle Progression.
PA1-964 was used in western blot to elucidate the roles of hRpn13 and Uch37 in cell cycle progression
|Randles L,Anchoori RK,Roden RB,Walters KJ||The Journal of biological chemistry (291:8773)||2016|
The capture proteasome assay: A method to measure proteasome activity in vitro.
PA1-964 was used in immunoprecipitation to develop the capture proteasome assay to assess proteasome activity in vitro using cell lysates.
|Vigneron N,Abi Habib J,Van den Eynde BJ||Analytical biochemistry (482:7)||2015|
Landscape of the PARKIN-dependent ubiquitylome in response to mitochondrial depolarization.
PA1-964 was used in western blot to use a proteomics approch to identify the range of PARKIN substrates during mitochondrial depolarization
|Sarraf SA,Raman M,Guarani-Pereira V,Sowa ME,Huttlin EL,Gygi SP,Harper JW||Nature (496:372)||2013|
Proteasomal non-catalytic subunit PSMD2 as a potential therapeutic target in association with various clinicopathologic features in lung adenocarcinomas.
PA1-964 was used in western blot to investigate the effect of PSMD2 knockdown in different lung cancer cell lines
|Matsuyama Y,Suzuki M,Arima C,Huang QM,Tomida S,Takeuchi T,Sugiyama R,Itoh Y,Yatabe Y,Goto H,Takahashi T||Molecular carcinogenesis (50:301)||2011|