Immunofluorescent analysis of Proteasome 20S X (green) showing staining in the cytoplasm and nucleus of A431 cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a Proteasome 20S X polyclonal antibody (Product # PA1-977) in 3% BSA-PBS at a dilution of 1:100 and incubated overnight at 4 °C in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a flourescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.
|Tested species reactivity||Bovine, Human, Mouse|
|Published species reactivity||Mouse, Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic Peptide: C W(243) I R V S S D N V A D L H D K Y S(259)|
|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 *|
|Western Blot (WB)||1:500-1:5000|
* 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 5 publications below|
PA1-977 detects proteasome 20S X from bovine, mouse and human samples.
PA1-977 has been successfully used in Western blot and ICC/IF procedures. By Western blot, this antibody detects an ~23 kDa protein representing proteasome 20S X.
The PA1-977 immunizing peptide corresponds to amino acid residues 243-259 from mouse proteasome 20S X. This sequence is 94% conserved in human and rat. PA1-977 immunizing peptide (Cat. # PEP-172) is available for use in neutralization and control procedures.
The proteasome is a multicatalytic proteinase complex with a highly ordered ring-shaped 20S core structure. The core structure is composed of 4 rings of 28 non-identical subunits; 2 rings are composed of 7 alpha subunits and 2 rings are composed of 7 beta subunits. Proteasomes are distributed throughout eukaryotic cells at a high concentration and cleave peptides in an ATP/ubiquitin-dependent process in a non-lysosomal pathway. An essential function of a modified proteasome, the immunoproteasome, is the processing of class I MHC peptides. This gene encodes a member of the proteasome B-type family, also known as the T1B family, that is a 20S core beta subunit in the proteasome. This catalytic subunit is not present in the immunoproteasome and is replaced by catalytic subunit 3i (proteasome beta 8 subunit). Multiple transcript variants encoding different isoforms have been found for this gene.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Denervation-Induced Activation of the Ubiquitin-Proteasome System Reduces Skeletal Muscle Quantity Not Quality.
PA1-977 was used in western blot investigate the role of the ubiquitin-proteasome system in skeletal muscle
|Baumann CW,Liu HM,Thompson LV||PloS one (11:null)||2016|
A selective inhibitor of the immunoproteasome subunit LMP2 induces apoptosis in PC-3 cells and suppresses tumour growth in nude mice.
PA1-977 was used in western blot to study the role of immunoproteasome subunit LMP2 in apoptosis of PC-3 cells and tumor growth in nude mice
|Wehenkel M,Ban JO,Ho YK,Carmony KC,Hong JT,Kim KB||British journal of cancer (107:53)||2012|
Persistent hijacking of brain proteasomes in HIV-associated dementia.
PA1-977 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.
PA1-977 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|
Resveratrol promotes clearance of Alzheimer's disease amyloid-beta peptides.
PA1-977 was used in western blot to demonstrate a proteasome-dependent anti-amyloidogenic activity of resveratrol.
|Marambaud P,Zhao H,Davies P||The Journal of biological chemistry (280:37377)||2005|