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PSMB3 Rabbit Polyclonal Antibody

Usd: 315

Specification

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Catalog# HA500458

PSMB3 Rabbit Polyclonal Antibody

Application

  • WB

  • IF-Cell

  • IHC-P

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

Overview

Product Name

PSMB3 Rabbit Polyclonal Antibody

Antibody Type

Rabbit Polyclonal Antibody

Immunogen

Recombinant full length protein within human PSMB3.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, IHC-P, FC

Target Molecular Weight

Predicted band size: 23 kDa

Positive Control

HeLa, Human skin tissue, Human breast tissue, mouse kidney tissue, rat kidney tissue, HeLa cell lysate, K-562 cell lysate, NIH/3T3 cell lysate, Mouse testis tissue lysate, Rat testis tissue lysate.

Conjugation

unconjugated

RRID

AB_3071543

Product Features

Form

Liquid

Concentration

2 mg/mL.(The concentration of this product may be batch-dependent)

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Immunogen affinity purified.

Application Dilution

  • WB

  • 1:5,000

  • IF-Cell

  • 1:200

  • IHC-P

  • 1:500-1:1,000

  • FC

  • 1:1,000

Target

Function

Non-catalytic component of the 20S core proteasome complex involved in the proteolytic degradation of most intracellular proteins. This complex plays numerous essential roles within the cell by associating with different regulatory particles. Associated with two 19S regulatory particles, forms the 26S proteasome and thus participates in the ATP-dependent degradation of ubiquitinated proteins. The 26S proteasome plays a key role in the maintenance of protein homeostasis by removing misfolded or damaged proteins that could impair cellular functions, and by removing proteins whose functions are no longer required. Associated with the PA200 or PA28, the 20S proteasome mediates ubiquitin-independent protein degradation. This type of proteolysis is required in several pathways including spermatogenesis (20S-PA200 complex) or generation of a subset of MHC class I-presented antigenic peptides (20S-PA28 complex).

Background References

1. Yano M., Koumoto Y., Kanesaki Y., Wu X., Kido H. 20S proteasome prevents aggregation of heat-denatured proteins without PA700 regulatory subcomplex like a molecular chaperone. Biomacromolecules 5:1465-1469 (2004)

2. Rut W., Drag M. Human 20S proteasome activity towards fluorogenic peptides of various chain lengths. Biol. Chem. 397:921-926 (2016)

Subcellular Location

Cytoplasm, Nucleus, Proteasome

UNIPROT

SwissProt: P49720 Human

SwissProt: Q9R1P1 Mouse

SwissProt: P40112 Rat

Synonyms

C10 II antibody

HC10 II antibody

proteasome (prosome, macropain) subunit, beta type 3 antibody

Proteasome beta 3 subunit antibody

Proteasome chain 13 antibody

proteasome component C10 II antibody

Proteasome component C10-II antibody

Proteasome subunit beta type-3 antibody

Proteasome theta chain antibody

PSB3_HUMAN antibody

Expand

Images

  • Immunocytochemistry analysis of HeLa cells labeling PSMB3 with Rabbit anti-PSMB3 antibody (<a href="/products/HA500458" style="font-weight: bold;text-decoration: underline;">HA500458</a>) at 1/200 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.05% Triton X-100 in PBS for 15 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-PSMB3 antibody (<a href="/products/HA500458" style="font-weight: bold;text-decoration: underline;">HA500458</a>) at 1/200 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Mouse anti-Beta tubulin Antibody (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>) was used as Microtubule Marker at 1/200 dilution and Goat Anti-Mouse IgG H&L (iFluor&trade; 647, <a href="/products/HA1127" style="font-weight: bold;text-decoration: underline;">HA1127</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of HeLa cells labeling PSMB3 with Rabbit anti-PSMB3 antibody (HA500458) at 1/200 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.05% Triton X-100 in PBS for 15 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-PSMB3 antibody (HA500458) at 1/200 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Mouse anti-Beta tubulin Antibody (M1305-2) was used as Microtubule Marker at 1/200 dilution and Goat Anti-Mouse IgG H&L (iFluor™ 647, HA1127) was used as the secondary antibody at 1/1,000 dilution.

  • Immunohistochemical analysis of paraffin-embedded Human skin tissue with Rabbit anti-PSMB3 antibody (<a href="/products/HA500458" style="font-weight: bold;text-decoration: underline;">HA500458</a>) at 1/600 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA500458" style="font-weight: bold;text-decoration: underline;">HA500458</a>) at 1/600 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded Human skin tissue with Rabbit anti-PSMB3 antibody (HA500458) at 1/600 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA500458) at 1/600 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded Human breast tissue with Rabbit anti-PSMB3 antibody (<a href="/products/HA500458" style="font-weight: bold;text-decoration: underline;">HA500458</a>) at 1/600 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA500458" style="font-weight: bold;text-decoration: underline;">HA500458</a>) at 1/600 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded Human breast tissue with Rabbit anti-PSMB3 antibody (HA500458) at 1/600 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA500458) at 1/600 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-PSMB3 antibody (<a href="/products/HA500458" style="font-weight: bold;text-decoration: underline;">HA500458</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA500458" style="font-weight: bold;text-decoration: underline;">HA500458</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-PSMB3 antibody (HA500458) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA500458) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-PSMB3 antibody (<a href="/products/HA500458" style="font-weight: bold;text-decoration: underline;">HA500458</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA500458" style="font-weight: bold;text-decoration: underline;">HA500458</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-PSMB3 antibody (HA500458) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA500458) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Flow cytometric analysis of HeLa cells labeling PSMB3.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/HA500458" style="font-weight: bold;text-decoration: underline;">HA500458</a>, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of HeLa cells labeling PSMB3.

    Cells were fixed and permeabilized. Then stained with the primary antibody (HA500458, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

  • Western blot analysis of PSMB3 on different lysates with Rabbit anti-PSMB3 antibody (<a href="/products/HA500458" style="font-weight: bold;text-decoration: underline;">HA500458</a>) at 1/5,000 dilution.<br /><br />Lane 1: HeLa cell lysate <br />Lane 2: K-562 cell lysate <br />Lane 3: NIH/3T3 cell lysate <br />Lane 4: Mouse testis tissue lysate <br />Lane 5: Rat testis tissue lysate <br /><br />Lysates/proteins at 30 µg/Lane.<br />Exposure time: 6 seconds; ECL: K1801<br /><br />Blocking: 5% NFDM/TBST, 1 hour at room temperature<br />Primary antibody: <a href="/products/HA500458" style="font-weight: bold;text-decoration: underline;">HA500458</a>, 1/5,000 in primary antibody dilution buffer (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>), overnight at 4 ℃<br />Secondary antibody: Goat anti-Rabbit IgG-HRP (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature<br /><br />Predicted band size: 23 kDa<br />Observed band size: 23 kDa

    Western blot analysis of PSMB3 on different lysates with Rabbit anti-PSMB3 antibody (HA500458) at 1/5,000 dilution.

    Lane 1: HeLa cell lysate
    Lane 2: K-562 cell lysate
    Lane 3: NIH/3T3 cell lysate
    Lane 4: Mouse testis tissue lysate
    Lane 5: Rat testis tissue lysate

    Lysates/proteins at 30 µg/Lane.
    Exposure time: 6 seconds; ECL: K1801

    Blocking: 5% NFDM/TBST, 1 hour at room temperature
    Primary antibody: HA500458, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 ℃
    Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

    Predicted band size: 23 kDa
    Observed band size: 23 kDa

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"