Western blot analysis was performed on whole cell extracts (30 µglysate) of MCF7 (Lane 1), PC-3 (Lane 2), Hep G2 (Lane 3), SK-OV-3 (Lane 4), HeLa (Lane 5), COS-7 (Lane 6), Jurkat (Lane 7), A-431 (Lane 8) and tissue extracts (30 µglysate) of Mouse Testis (Lane 9) and Rat Testis (Lane 10). The blot was probed with Anti-PSMD1 Rabbit Polyclonal Antibody (Product # PA1-973, 1 µg/ml) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/ml, 1:2500 dilution). A 106 kDa band corresponding to PSMD1 was observed across the cell lines and tissues tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 10 % Bis-Tris gel (Product # NP0302BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with overnight wet transfer system. The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
|Tested species reactivity||Bovine, Human, Mouse, Non-human primate, Rat|
|Published species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic Peptide: A(929) A H G P K I E E E E Q E P E P P E P F E Y I D D(953)|
|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|
|Immunohistochemistry (IHC)||15 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.
PA1-973 detects proteasome 19S subunit S1 from from human cells.
PA1-973 has been successfully used in Western blot and immunohistochemistry procedures. By Western blot, this antibody detects an ~105 kDa protein representing proteasome 19S subunit S1 from HeLa cell extract.
PA1-973 immunizing peptide corresponds to amino acid residues 929-953 from human proteasome 19S subunit S1 protein.
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.
Glucosamine induces cell death via proteasome inhibition in human ALVA41 prostate cancer cell.
PA1-973 was used in western blot to investigate the mechanism for glucosamine-induced cell apoptosis in cancer cells
|Liu BQ,Meng X,Li C,Gao YY,Li N,Niu XF,Guan Y,Wang HQ||Experimental and molecular medicine (43:487)||2011|
Persistent hijacking of brain proteasomes in HIV-associated dementia.
PA1-973 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|
Reduced ubiquitin C-terminal hydrolase-1 expression levels in dementia with Lewy bodies.
PA1-973 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|
Neuropathology and pathogenesis of encephalitis following amyloid-beta immunization in Alzheimer's disease.
PA1-973 was used in immunohistochemistry to investigate the cause of meningoencephalitis after immunization with amyloid-beta peptide in Alzheimer disease
|Ferrer I,Boada Rovira M,Sánchez Guerra ML,Rey MJ,Costa-Jussá F||Brain pathology (Zurich, Switzerland) (14:11)||2004|