Revision 1

#12446Store at -20C

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IP

REACTIVITY:

H M R Mk

SENSITIVITY:

Endogenous

MW (kDa):

28

Source/Isotype:

Rabbit IgG

UniProt ID:

#P25788

Entrez-Gene Id:

5684

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Specificity / Sensitivity

PSMA3 (D4Y9O) Rabbit mAb recognizes endogenous levels of total PSMA3 protein. This antibody does not cross-react with other α subunits of the 20S proteasome. This antibody cross-reacts with a 140 kDa protein of unknown identity in lysates from certain cell lines.

Species Reactivity:

Human, Mouse, Rat, Monkey

Species predicted to react based on 100% sequence homology

Dog, Pig, Horse

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Arg100 of human PSMA3 protein.

Background

The 26S proteasome is a highly abundant proteolytic complex involved in the degradation of ubiquitinated substrate proteins. It consists largely of two sub-complexes, the 20S catalytic core particle (CP) and the 19S/PA700 regulatory particle (RP) that can cap either end of the CP. The CP consists of two stacked heteroheptameric β-rings (β1-7) that contain three catalytic β-subunits and are flanked on either side by two heteroheptameric α-rings (α1-7). The RP includes a base and a lid, each having multiple subunits. The base, in part, is composed of a heterohexameric ring of ATPase subunits belonging to the AAA (ATPases Associated with diverse cellular Activities) family. The ATPase subunits function to unfold the substrate and open the gate formed by the α-subunits, thus exposing the unfolded substrate to the catalytic β-subunits. The lid consists of ubiquitin receptors and DUBs that function in recruitment of ubiquitinated substrates and modification of ubiquitin chain topology (1,2). Other modulators of proteasome activity, such as PA28/11S REG, can also bind to the end of the 20S CP and activate it (1,2).
PSMA3 (HC8/α7) is an α subunit of the 20S proteasome. PSMA3 lacks any known proteolytic activity but may have an important role in the intiation of ring formation and 20S proteasome assembly (3). Research studies have demonstrated that PSMA3 serves as a receptor within the proteasome, recruiting substrates that are destined for degradation in a ubiquitination-independent manner (4,5).

  1. Finley, D. (2009) Annu Rev Biochem 78, 477-513.
  2. Lee, M.J. et al. (2011) Mol Cell Proteomics 10, R110.003871.
  3. Gerards, W.L. et al. (1998) J Mol Biol 275, 113-21.
  4. Touitou, R. et al. (2001) EMBO J 20, 2367-75.
  5. Sdek, P. et al. (2005) Mol Cell 20, 699-708.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: 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.

Applications Key

WB: Western Blotting IP: Immunoprecipitation

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Revision 1
#12446

PSMA3 (D4Y9O) Rabbit mAb

Western Blotting Image 1: PSMA3 (D4Y9O) Rabbit mAb Expand Image
Western blot analysis of extracts from various cell lines using PSMA3 (D4Y9O) Rabbit mAb.
Western Blotting Image 2: PSMA3 (D4Y9O) Rabbit mAb Expand Image
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with constructs expressing Myc/DDK-tagged full-length human PSMA1 (hPSMA1-Myc/DDK; +), full-length human PSMA2 (hPSMA2-Myc/DDK; +), full-length human PSMA3 (hPSMA3-Myc/DDK; +), full-length human PSMA4 (hPSMA4-Myc/DDK; +), full-length human PSMA6 (hPSMA6-Myc/DDK; +), or full-length human PSMA7 (hPSMA7-Myc/DDK; +), using PSMA3 (D4Y9O) Rabbit mAb (upper) or DYKDDDDK Tag Antibody #2368 (lower).
Immunoprecipitation Image 1: PSMA3 (D4Y9O) Rabbit mAb Expand Image
Immunoprecipitation of PSMA3 from 293T cell extracts, using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or PSMA3 (D4Y9O) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using PSMA3 (D4Y9O) Rabbit mAb.