Recombinant protein within Human PSMD14 aa 92-230 / 310.
Species Reactivity
Human, Rat, Mouse
Validated Applications
WB, IHC-P
Target Molecular Weight
Predicted band size: 35 kDa
Positive Control
Human skin tissue lysate, Rat bone marrow lysate, human liver cancer tissue, human breast tissue, human pancreas tissue, mouse kidney tissue, mouse heart tissue.
As the degradation machinery that is responsible for ~70% of intracellular proteolysis, the proteasome complex (26S proteasome) plays critical roles in maintaining the homeostasis of the cellular proteome. Misfolded proteins and damaged protein need to be continuously removed to recycle amino acids for new synthesis; in addition, some key regulatory proteins fulfil their biological functions via selective degradation; furthermore, proteins are digested into peptides for MHC class I antigen presentation. To meet such complicated demands in biological processes via spatial and temporal proteolysis, protein substrates have to be recognized, recruited, and eventually hydrolyzed in a controlled fashion. Thus, the 19S regulatory particle has a series of important capabilities to address these functional challenges. To recognize proteins as designated substrates, the 19S complex has subunits that are capable of recognizing proteins with a special degradative tag, ubiquitination. It also has subunits that can bind to nucleotides (e.g., ATPs) in order to facilitate the association between the 19S and 20S particles, as well as to cause conformational changes to the alpha subunit C-terminals that form the substate entrance of the 20S complex. Rpn11 drives metalloprotease activity to hydrolyze the ubiquitin molecules from the poly-ubiquitin chain before protein substrates are unfolded and degraded.
Background References
1. Spataro V et al. Resistance to diverse drugs and ultraviolet light conferred by overexpression of a novel human 26 S proteasome subunit. J. Biol. Chem. 272:30470-30475 (1997).
2. Kanayama HO et al. Demonstration that a human 26S proteolytic complex consists of a proteasome and multiple associated protein components and hydrolyzes ATP and ubiquitin-ligated proteins by closely linked mechanisms. Eur. J. Biochem. 206:567-578 (1992).
Sequence Similarity
Belongs to the peptidase M67A family. PSMD14 subfamily.
Tissue Specificity
Widely expressed. Highest levels in heart and skeletal muscle.
Subcellular Location
Cytosol. Extracellular region or secreted. Nucleus.
Immunohistochemical analysis of paraffin-embedded human liver cancer tissue using anti-PSMD14 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7109-85, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Immunohistochemical analysis of paraffin-embedded human breast tissue using anti-PSMD14 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7109-85, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Immunohistochemical analysis of paraffin-embedded human pancreas tissue using anti-PSMD14 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7109-85, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Immunohistochemical analysis of paraffin-embedded mouse kidney tissue using anti-PSMD14 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7109-85, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Immunohistochemical analysis of paraffin-embedded mouse heart tissue using anti-PSMD14 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7109-85, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Western blot analysis of PSMD14 on different lysates with Rabbit anti-PSMD14 antibody (ET7109-85) at 1/1,000 dilution.
Lane 1: Rat bone marrow tissue lysate
Lysates/proteins at 40 µg/Lane. Exposure time: 40 seconds; ECL: K1801
Blocking: 5% NFDM/TBST, 1 hour at room temperature Primary antibody: ET7109-85, 1/1,000 in primary antibody dilution buffer (K1803), overnight at 4 ℃ Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature
Predicted band size: 35 kDa Observed band size: 35 kDa
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