Invitrogen

PSMB9 Monoclonal Antibody (OTI1C4)

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Protocols

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PSMB9 Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of Proteasome subunit beta type-9 was performed using 70% confluent log phase SK-O-V3 cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.1% Triton™ X-100 for 45 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with PSMB9 Monoclonal Antibody (OTI1C4) (Product # MA5-25887) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175), (1:2000 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300 dilution). Panel d represents the merged image showing cytoplasmic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

PSMB9 Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemistry was performed on paraffin-embedded adenocarcinoma of human endometrium tissue. To expose target proteins, 10mM citric buffer, pH6.0, 100°C for 10min was used. Following antigen retrieval, tissues were probed with a PSMB9 monoclonal antibody (Product # MA5-25887).

PSMB9 Antibody in Western Blot (WB) — Western blot was performed using Anti-PSMB9 Monoclonal Antibody (OTI1C4) (Product # MA5-25887) and a 20 kDa band corresponding to Proteasome subunit beta type-9 was observed across the cell lines and tissues tested. Whole cell extracts (30 µg lysate) of HEL 92.1.7 (Lane 1), Reh (Lane 2), SK-O-V3 (Lane 3), Raji (Lane 4) and Mouse Spleen (Lane 5) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # LC2001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177,1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescentdetection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Product Details

MA5-25887

Applications	Tested Dilution	Publications
Western Blot (WB)	1:1,000-1:2,000	-
Immunohistochemistry (Paraffin) (IHC (P))	1:150	-
Immunocytochemistry (ICC/IF)	1:100	-
Flow Cytometry (Flow)	1:100	-

Product Specifications
Species Reactivity	Human, Mouse
Host/Isotype	Mouse / IgG2b
Class	Monoclonal
Type	Antibody
Clone	OTI1C4
Immunogen	Human recombinant protein fragment corresponding to amino acids 21-219 of PSMB9 produced in HEK293T cell View immunogen
Conjugate	Unconjugated
Form	Liquid
Concentration	1 mg/mL
Purification	Affinity Chromatography
Storage buffer	PBS, pH 7.3, with 1% BSA, 50% glycerol
Contains	0.02% sodium azide
Storage conditions	-20° C, Avoid Freeze/Thaw Cycles
RRID	AB_2725132

Target Information

The proteasome is a multicatalytic proteinase complex with a highly ordered ring-shaped 20S core structure. The core structure 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. 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 a member of the proteasome B-type family, also known as the T1B family, that is a 20S core beta subunit. This gene is located in the class II region of the MHC in the immunoproteasome. Proteolytic processing is required to generate a mature subunit.

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

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Bioinformatics

Protein Aliases: large multifunctional peptidase 2; large multifunctional protease 2; LMP-2d; Low molecular mass protein 2; Macropain chain 7; MGC70470; Multicatalytic endopeptidase complex chain 7; proteasome (prosome, macropain) subunit, beta type, 9 (large multifunctional peptidase 2); proteasome catalytic subunit 1i; Proteasome chain 7; proteasome subunit beta 6i; proteasome subunit beta type 9; Proteasome subunit beta type-9; Proteasome subunit beta-1i; proteasome-related gene 2; proteosome (prosome, macropain) subunit, beta type 9 (large multifunctional peptidase 2); proteosome beta 9 subunit; Really interesting new gene 12 protein

Gene Aliases: beta1i; Lmp-2; LMP2; PSMB6i; PSMB9; RING12

UniProt ID: (Human) P28065, (Mouse) P28076

Entrez Gene ID: (Human) 5698, (Mouse) 16912

Molecular Function: protease