Immunofluorescence analysis of PSMD7 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 PSMD7 Rabbit Polyclonal Antibody (PA11963) 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 cytosolic localization. Panel e shows the control without primary antibody. The images were captured at 60X magnification.
|Tested species reactivity||Hamster, Human, Mouse, Rat|
|Published species reactivity||Mouse, Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to residues A(55) V P F D E D D K D D S(66) C of human 19S subunit S12.|
|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 *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Western Blot (WB)||0.5-2 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|
PA1-1963 detects proteasome 19S subunit S12 from human, rat, mouse and hamster cell lysates.
PA1-1963 has been successfully used in Western blot procedures. By Western blot, this antibody detects an ~37 kDa protein representing proteasome 19S subunit S12 from human, rat, mouse, and hamster cell extracts. An additional unknown band is detected at ~48 kDa in some cell lysates.
PA1-1963 immunogen is a synthetic peptide corresponding to residues A(55) V P F D E D D K D D S(66) C of human 19S subunit S12. The immunizing peptide sequence is 100% conserved in Danio rerio and 91% conserved in Drosphila, C. elegans and Anopheles. PA1-1963 immunizing peptide (Cat. # PEP-211) is available for use in neutralization and control experiments.
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 signaling 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.
Interferon ß induces clearance of mutant ataxin 7 and improves locomotion in SCA7 knock-in mice.
PA1-1963 was used in western blot to study the ability of IFNbeta to induce PML-dependent ataxin 7 clearance and improve motor function in a murine model of spinocerebellar ataxia 7
|Chort A,Alves S,Marinello M,Dufresnois B,Dornbierer JG,Tesson C,Latouche M,Baker DP,Barkats M,El Hachimi KH,Ruberg M,Janer A,Stevanin G,Brice A,Sittler A||Brain : a journal of neurology (136:1732)||2013|
Persistent hijacking of brain proteasomes in HIV-associated dementia.
PA1-1963 was used in western blot to study the effect of HIV infection on brain proteasomes and its role in dementia
|Nguyen TP,Soukup VM,Gelman BB||The American journal of pathology (176:893)||2010|