Invitrogen

PSMB5 Polyclonal Antibody

Advanced Verification

This Antibody was verified by Knockdown to ensure that the antibody binds to the antigen stated.

5 Published Figures

9 References

View all (6) PSMB5 antibodies

Datasheet

Protocols

Questions & Answers

Tech Support

Datasheet

Protocols

Questions & Answers

Tech Support

Product Image Gallery

PSMB5 Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of PSMB5 was performed using 70% confluent log phase DU 145 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with PSMB5 Polyclonal Antibody (Product # PA1-977) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (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). Panel d represents the merged image showing nuclear localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

PSMB5 Antibody in Western Blot (WB) — Western blot was performed using Anti-PSMB5 Polyclonal Antibody (Product # PA1-977) and a 22kDa band corresponding to PSMB5 was observed across cell lines and tissue tested. Nuclear enriched extracts (30 µg lysate) of A-431 (Lane 1), DU 145 (Lane 2), MCF7 (Lane 3), NIH/3T3 (Lane 4), Mouse Kidney (Lane 5), Mouse Liver (Lane 6), Rat Liver (Lane 7), Mouse Heart (Lane 8) were electrophoresed using NuPAGE™ 12% Bis-Tris Protein Gel (Product # NP0342BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23002) 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-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:6000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). Few uncharacterized bands were observed between 30-50 kDa.

PSMB5 Antibody — Figure 5 ( a ) Immunoblots of proteasome catalytic subunits in H727 cells transfected with siRNA targeting beta5, beta2 or beta5i. ( b ) The catalytic activity of beta5 subunit was decreased in H727 cells transfected with siRNA targeting beta5 compared with H727 cells transfected with scrambled siRNA. ( c ) Effects of siRNA knockdown of beta5, beta5i, or beta2 on Cfz sensitivity (IC 50 values) in H23 and H727 cells. Data are shown as mean +- SD derived from a single non-linear regression based on n = 3 replicates per compound per concentration. ( d ) Heat map showing proteasome catalytic subunit activity profiles of 6 PI-naive patient MM samples purchased from Conversant Biologics and AllCells (left). The numbers represent hydrolysis rates of respective substrates (RFU/min, mean values derived from three technical replicates) and were converted to color format and clustered by using the program ""R"" ( http://www.R-project.org ) (left panel). Carfilzomib (Cfz) cell viability of the same 6 patient MM cells was measured via CellTiter-Glo Luminescent Cell Viability Assay (right panel). {KD}

Product Details

PA1-977

Applications	Tested Dilution	Publications
Western Blot (WB)	1:500-1:5,000	View 8 publications 8 publications
Immunocytochemistry (ICC/IF)	1:10-1:200	View 1 publication 1 publication

Product Specifications
Species Reactivity	Bovine, Human, Mouse
Published species	Human, Mouse
Host/Isotype	Rabbit / IgG
Class	Polyclonal
Type	Antibody
Immunogen	Synthetic Peptide: C W(243) I R V S S D N V A D L H D K Y S(259)
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_2172052

Product Specific Information

PA1-977 detects proteasome 20S X from bovine, mouse and human samples.

PA1-977 has been successfully used in Western blot and ICC/IF procedures. By Western blot, this antibody detects an ~23 kDa protein representing proteasome 20S X.

The PA1-977 immunizing peptide corresponds to amino acid residues 243-259 from mouse proteasome 20S X. This sequence is 94% conserved in human and rat. PA1-977 immunizing peptide (Cat. # PEP-172) is available for use in neutralization and control procedures.

Target Information

Cyclic nucleotide phosphodiesterases.

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

Product References

H727 cells are inherently resistant to the proteasome inhibitor carfilzomib, yet require proteasome activity for cell survival and growth.

Scientific reports

Lee MJ,Miller Z,Park JE,Bhattarai D,Lee W,Kim KB

PA1-977 was used in Western Blotting to assess the use of H727 human bronchial carcinoid cells as a model for de novo carfilzomib resistance.

Mon Mar 11 00:00:00 EDT 2019

Skeletal muscle denervation investigations: selecting an experimental control wisely.

American journal of physiology. Cell physiology

Liu H,Thompson LV

PA1-977 was used in Western Blotting to investigate whether skeletal denervation unilaterally alters the proteolytic system in the contralateral-innervated muscles.

Fri Mar 01 00:00:00 EST 2019

Novel Potent Capsid Assembly Modulators Regulate Multiple Steps of the Hepatitis B Virus Life Cycle.

Antimicrobial agents and chemotherapy

Lahlali T,Berke JM,Vergauwen K,Foca A,Vandyck K,Pauwels F,Zoulim F,Durantel D

PA1-977 was used in Immunocytochemistry-immunoflourescence to demonstrate that some capsid assembly modulators could interfere with multiple functions of Hepatitis B virus core protein in the viral life cycle.

Mon Oct 01 00:00:00 EDT 2018

Induction of the Immunoproteasome Subunit Lmp7 Links Proteostasis and Immunity in α-Synuclein Aggregation Disorders.

EBioMedicine

Ugras S,Daniels MJ,Fazelinia H,Gould NS,Yocum AK,Luk KC,Luna E,Ding H,McKennan C,Seeholzer S,Martinez D,Evans P,Brown D,Duda JE,Ischiropoulos H

PA1-977 was used in Western Blotting to examine the role of the Lmp7 subunit of the immunoproteasome in the degradation of α-synuclein aggregates.

Tue May 01 00:00:00 EDT 2018

Denervation-Induced Activation of the Ubiquitin-Proteasome System Reduces Skeletal Muscle Quantity Not Quality.

PloS one

Baumann CW,Liu HM,Thompson LV

PA1-977 was used in western blot investigate the role of the ubiquitin-proteasome system in skeletal muscle

Fri Jul 28 00:00:00 EDT 2017

A selective inhibitor of the immunoproteasome subunit LMP2 induces apoptosis in PC-3 cells and suppresses tumour growth in nude mice.

British journal of cancer

Wehenkel M,Ban JO,Ho YK,Carmony KC,Hong JT,Kim KB

PA1-977 was used in western blot to study the role of immunoproteasome subunit LMP2 in apoptosis of PC-3 cells and tumor growth in nude mice

Tue Jun 26 00:00:00 EDT 2012

Persistent hijacking of brain proteasomes in HIV-associated dementia.

The American journal of pathology

Nguyen TP,Soukup VM,Gelman BB

PA1-977 was used in western blot to study the effect of HIV infection on brain proteasomes and its role in dementia

Mon Feb 01 00:00:00 EST 2010

Reduced ubiquitin C-terminal hydrolase-1 expression levels in dementia with Lewy bodies.

Neurobiology of disease

Barrachina M,Castaño E,Dalfó E,Maes T,Buesa C,Ferrer I

PA1-977 was used in western blot to investigate the changes of UCHL-1 in samples from dementia patients with Lewy bodies

Mon May 01 00:00:00 EDT 2006

Resveratrol promotes clearance of Alzheimer's disease amyloid-beta peptides.

The Journal of biological chemistry

Marambaud P,Zhao H,Davies P

PA1-977 was used in western blot to demonstrate a proteasome-dependent anti-amyloidogenic activity of resveratrol.

Fri Nov 11 00:00:00 EST 2005

Bioinformatics

Protein Aliases: Macropain epsilon chain; MGC118075; Multicatalytic endopeptidase complex epsilon chain; proteasome (prosome, macropain) subunit, beta type, 5; proteasome beta 5 subunit; proteasome beta type subunit 5; proteasome catalytic subunit 3; Proteasome chain 6; Proteasome epsilon chain; proteasome subunit beta 5; Proteasome subunit beta type-5; Proteasome subunit MB1; Proteasome subunit X; proteasome subunit, beta type, 5; PSX large multifunctional protease X; testicular tissue protein Li 153

Gene Aliases: LMPX; MB1; PSMB5; X

UniProt ID: (Human) P28074, (Bovine) Q32KL2, (Mouse) O55234

Entrez Gene ID: (Human) 5693, (Bovine) 534640, (Mouse) 19173

Molecular Function: protease