Western blot analysis of extracts from HeLa and SW620 cells, untreated (-) or treated with Human Interferon-γ (hIFN-γ) #8901 (100 ng/ml, 48 hr; +), using PSMB10/MECL-1 Antibody (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western blot analysis of extracts from HeLa and B16-F10 cells, untreated (-) or treated with Interferon-γ (IFN-γ) (100 ng/ml, 48 hr; +), using PSMB10/MECL-1 Antibody (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected (+) with constructs expressing Myc/DDK-tagged full-length human PSMB10 protein (hPSMB10-Myc/DDK) and Myc/DDK-tagged full-length human PSMB7 protein (hPSMB7-Myc/DDK), using PSMB10/MECL-1 Antibody (upper), DYKDDDDK Tag Antibody #2368 (middle), and β-Actin (D6A8) Rabbit mAb #8457 (lower).
|MW (kDa)||25, 29|
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Store at –20°C. Do not aliquot the antibody.
For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
NOTE: Please refer to primary antibody product webpage for recommended antibody dilution.
From sample preparation to detection, the reagents you need for your Western Blot are now in one convenient kit: #12957 Western Blotting Application Solutions Kit
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
Load 20 µl onto SDS-PAGE gel (10 cm x 10 cm).
NOTE: Volumes are for 10 cm x 10 cm (100 cm2) of membrane; for different sized membranes, adjust volumes accordingly.
* Avoid repeated exposure to skin.
posted June 2005
revised June 2020
Protocol Id: 10
PSMB10/MECL-1 Antibody recognizes endogenous levels of total PSMB10/MECL-1 protein. This antibody does not cross-react with PSMB7/Z protein.
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human PSMB10/MECL-1 protein. Antibodies are purified by protein A and peptide affinity chromatography.
The 20S proteasome is the major proteolytic enzyme complex involved in intracellular protein degradation. It consists of four stacked rings, each with seven distinct subunits. The two outer layers are identical rings composed of α subunits (called PSMAs), and the two inner layers are identical rings composed of β subunits. While the catalytic sites are located on the β rings (1-3), the α subunits are important for assembly and as binding sites for regulatory proteins (4). Seven different α and ten different β proteasome genes have been identified in mammals (5). PA700, PA28, and PA200 are three major protein complexes that function as activators of the 20S proteasome. PA700 binds polyubiquitin with high affinity and associates with the 20S proteasome to form the 26S proteasome, which preferentially degrades polyubiquitinated proteins (1-3). The proteasome has a broad substrate spectrum that includes cell cycle regulators, signaling molecules, tumor suppressors, and transcription factors. By controlling the degradation of these intracellular proteins, the proteasome functions in cell cycle regulation, cancer development, immune responses, protein folding, and disease progression (6-9).
Proteasome subunit β type-10 (PSMB10, LMP10, MECL-1) is expressed as a proenzyme that is autocleaved to form a mature immunoproteasome core particle subunit (10). Like other immunoproteasome subunits, PSMB10/MECL-1 expression is induced by IFN-γ and subsequent replacement of constitutively expressed PSMB7/Z within the 20S proteasome proteolytic core particle facilitates processing and presentation of MHC class I-restricted peptide antigens on the cell surface (11-13).
Explore pathways + proteins related to this product.
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