Product Pathways - Protein Stability
PSMB8/LMP7 (1A5) Mouse mAb #13726
|13726S||100 µl (10 western blots)||---||In Stock||---|
|13726||carrier free and custom formulation / quantity||email request|
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|W||1:1000||Human, Rat||Endogenous||23, 28||Mouse IgG1|
Species cross-reactivity is determined by western blot.
Applications Key: W=Western Blotting, IF-IC=Immunofluorescence (Immunocytochemistry), F=Flow Cytometry
* Product-specific protocol.
Specificity / Sensitivity
PSMB8/LMP7 (1A5) Mouse mAb recognizes endogenous levels of total PSMB8/LMP7 protein. This antibody recognizes both 28 kDa precursor and 23 kDa mature forms of PSMB8/LMP7 and does not cross-react with PSMB5 protein.
Source / Purification
Monoclonal antibody is produced by immunizing animals with recombinant protein encompassing the full-length of human PSMB8/LMP7 protein.
Western blot analysis of extracts from various cell lines using PSMB8/LMP7 (1A5) Mouse mAb (upper) and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). (The T2 (13) and RKO (8) cell lines are both deficient in PSMB8/LMP8 protein expression).
Western blot analysis of extracts from HeLa and SW620 cells, untreated (-) or treated with Human Interferon-γ (hIFN-γ) #8901 (100 ng/ml, 72 hr; +), using PSMB8/LMP7 (1A5) Mouse mAb (upper) and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with constructs expressing Myc/DDK-tagged full-length human PSMB5 (hPSMB5-Myc/DDK; +) or Myc/DDK-tagged full-length human PSMB8 (hPSMB8-Myc/DDK; +), using PSMB8/LMP7 (1A5) Mouse mAb (upper) and DYKDDDDK Tag Antibody #2368 (lower).
Flow cytometric analysis of Raji cells using PMSB8/LMP7 (1A5) Mouse mAb (blue) compared to Mouse (G3A1) mAb IgG1 Isotype Control #5415 (Red). Anti-mouse IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4408 was used as a secondary antibody.
The 26S proteasome is a highly abundant, ~2 MDa complex that serves as the proteolytic arm of the ubiquitin-proteasome system. It consists largely of two sub-complexes, the 19S regulatory particle (RP) and the 20S catalytic core particle (CP); in many cases two RPs cap either end of a CP. The CP is made of two stacked β-rings that contain the catalytic sites, each of which is made of seven subunits (β1-7), flanked on either side by two α-rings, which are also made of seven subunits each (α1-7). Thus, the structure of the 20S CP is α1-7β1-7β1-7α1-7. The RP includes a base and a lid. The base, in part, is composed of a hexametric ring of ATPases that function to unfold the substrate and open the gate of the interlacing amino-terminal segments of the α-subunits, thus allowing entry of the unfolded substrate into the catalytic chamber. The lid is predominantly involved in specific recognition of the ubiquitin signal (1). In addition to the 19S cap, other proteins and complexes, such as proteasome activator 28 (PA28/11S), bind to the end of the 20S cylinder and activate it by facilitating opening of the gate. Furthermore, proteasome-associated DUBs and E3s can remodel substrate-anchored polyubiquitin chains, which may modulate their susceptibility to degradation (2).
Constitutively expressed core particle subunits PSMB5, PSMB7, and PSMB6 provide chymotrypsin-like, trypsin-like, and caspase-like activities, respectively (3). In immune cells involved in antigen presentation, these subunits are replaced by highly homologous, induced β-subunits to form the immunoproteasome (4,5).
Proteasome subunit beta type-8 (PSMB8, LMP7) is expressed as a proenzyme that is cleaved to form the mature PSMB8 (LMP7) immunoproteasome core particle subunit (6). Interferon-γ induces expression of PSMB8, which functionally replaces the PSMB5 core particle subunit in immunoproteasome processing of MHC class I-restricted peptide antigens (7). Research studies suggest that reduced PSMB8 expression or expression of the non-functional LMP7-E1 isoform may impair immunoproteasome assembly, and that PSMB8 deficiency results in reduced MHC class I molecule expression (8-10). Inhibition of PSMB8 in murine rheumatoid arthritis models attenuates disease indicators, suggesting that PSMB8 is a potential therapeutic target in the treatment of some proinflammatory autoimmune diseases (11). Mutations in the corresponding PSMB8 gene can cause an autoinflammatory syndrome known as CANDLE Syndrome (12).
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- De, M. et al. (2003) J Biol Chem 278, 6153-9.
- Fehling, H.J. et al. (1994) Science 265, 1234-7.
- Muchamuel, T. et al. (2009) Nat Med 15, 781-7.
- Liu, Y. et al. (2012) Arthritis Rheum 64, 895-907.
- Salter, R.D. et al. (1985) Immunogenetics 21, 235-46.
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For Research Use Only. Not For Use In Diagnostic Procedures.
Tween® is a registered trademark of ICI Americas, Inc.
Alexa Fluor® is a registered trademark of Molecular Probes, Inc.
Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.
This antibody is developed, validated, and produced by CST using in part technology under license (granting certain rights including those under U.S. Patents No. 5,675,063 and 7,429,487) from Epitomics, Inc.