Invitrogen

PSMD3 Polyclonal Antibody

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PSMD3 Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescent analysis of Proteasome 19S Subunit S3 (green) showing staining in the cytoplasm and nucleus of NIH-3T3 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 19S Subunit S3 polyclonal antibody (Product # PA1-974) 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 fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.

PSMD3 Antibody in Flow Cytometry (Flow) — Flow cytometry analysis of Proteasome 19S was done on HeLa cells. Cells were fixed, permeabilized and stained with a Proteasome 19S rabbit polyclonal antibody (Product # PA1-974, pink histogram) or a rabbit IgG isotype control (Product # MA5-16384, black histogram) at a dilution of 10 µg/mL. After incubation for 1 hour on ice, the cells were labeled with a Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 647 conjugate (Product # A27040) at a dilution of 1:50 for 1 hour on ice. A representative 10,000 cells were acquired and analyzed for each sample.

Product Details

PA1-974

Applications	Tested Dilution	Publications
Western Blot (WB)	1.0 µg/mL	-
Immunocytochemistry (ICC/IF)	1:50-1:500	-
Flow Cytometry (Flow)	1-20 µg/mL	-
Miscellaneous PubMed (Misc)	-	View 1 publication 1 publication

Product Specifications
Species Reactivity	Human, Mouse
Published species	Not Applicable
Host/Isotype	Rabbit / IgG
Class	Polyclonal
Type	Antibody
Immunogen	Synthetic Peptide: E(513) R E Q Q D L E F A K E M A E D D D D S F P(534)
Conjugate	Unconjugated
Form	Liquid
Concentration	1 mg/mL
Purification	Antigen affinity chromatography
Storage buffer	PBS with 1mg/mL BSA
Contains	0.05% sodium azide
Storage conditions	-20° C, Avoid Freeze/Thaw Cycles
RRID	AB_2237636

Product Specific Information

PA1-974 detects proteasome 19S subunit S3 from human and mouse cells.

PA1-974 has been successfully used in ICC/IF, FACS, and Western blot procedures. By Western blot, this antibody detects an ~60 kDa protein representing proteasome 19S subunit S3 from HeLa cell extract.

For Western blot procedures it is necessary to transfer the protein to membrane at pH 11.

PA1-974 immunizing peptide corresponds to amino acid residues 513-534 from human proteasome 19S subunit S3 protein.

Target Information

The 26S proteasome is a multicatalytic proteinase complex with a highly ordered structure composed of 2 complexes, a 20S core and a 19S regulator. The 20S core 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. The 19S regulator is composed of a base, which contains 6 ATPase subunits and 2 non-ATPase subunits, and a lid, which contains up to 10 non-ATPase 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 one of the non-ATPase subunits of the 19S regulator lid.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Product References

The cyclophilin-like domain of Ran-binding protein-2 modulates selectively the activity of the ubiquitin-proteasome system and protein biogenesis.

The Journal of biological chemistry

Yi H,Friedman JL,Ferreira PA

PA1-974 was used in western blot to investigate the role of proteasome subunits S10B during the proteolysis of intracellular proteins regulated by the ubiquitin-proteasome system.

Fri Nov 30 00:00:00 EST 2007

Bioinformatics

Protein Aliases: 26S proteasome non-ATPase regulatory subunit 3; 26S proteasome regulatory subunit RPN3; 26S proteasome regulatory subunit S3; proteasome (prosome, macropain) 26S subunit, non-ATPase, 3; Proteasome subunit p58; tissue specific transplantation antigen 2; tissue specific transplantation antigen P91A; Transplantation antigen P91A; Tum-P91A antigen

Gene Aliases: AI255837; AntP91a; D11Bwg1349e; P58; P91a; Psd3; PSMD3; RPN3; S3; TSTA2; Tstap91a

UniProt ID: (Human) O43242, (Mouse) P14685

Entrez Gene ID: (Human) 5709, (Mouse) 22123

Molecular Function: protease