Skip to content

Proteasome 20S LMP7 Recombinant Rabbit Monoclonal Antibody [JB54-32]

Usd: 385

Specification

Bulk Order Quote | Customization Request

Catalog# ET7107-36

Proteasome 20S LMP7 Recombinant Rabbit Monoclonal Antibody [JB54-32]

Application

  • WB

  • IF-Cell

  • IF-Tissue

  • IHC-P

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

Proteasome 20S LMP7 Recombinant Rabbit Monoclonal Antibody [JB54-32]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within Human Proteasome 20S LMP7 aa 160-276 / 276.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, IF-Tissue, IHC-P, FC

Molecular Weight

Predicted band size: 30 kDa

Positive Control

U-937 cell lysate, A431 cell lysate, U-937, HUVEC, human kidney tissue, mouse kidney tissue, mouse small intestine tissue, rat kidney tissue.

Conjugation

unconjugated

Clone Number

JB54-32

RRID

AB_3070748

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:500-1:1,000

  • IF-Cell

  • 1:100

  • IHC-P

  • 1:200-1:1,000

  • FC

  • 1:1,000

Target

Function

Proteasome subunit beta type-8 as known as 20S proteasome subunit beta-5i is a protein that in humans is encoded by the PSMB8 gene. This protein is one of the 17 essential subunits (alpha subunits 1–7, constitutive beta subunits 1–7, and inducible subunits including beta1i, beta2i, beta5i) that contributes to the complete assembly of 20S proteasome complex. In particular, proteasome subunit beta type-5, along with other beta subunits, assemble into two heptameric rings and subsequently a proteolytic chamber for substrate degradation. This protein contains "Chymotrypsin-like" activity and is capable of cleaving after large hydrophobic residues of peptide. The eukaryotic proteasome recognized degradable proteins, including damaged proteins for protein quality control purpose or key regulatory protein components for dynamic biological processes. The constitutive subunit beta1, beta2, and beta 5 (systematic nomenclature) can be replaced by their inducible counterparts beta1i, 2i, and 5i when cells are under the treatment of interferon-γ. The resulting proteasome complex becomes the so-called immunoproteasome. An essential function of the modified proteasome complex, the immunoproteasome, is the processing of numerous MHC class-I restricted T cell epitopes.

Background References

1. Akiyama K et al. Replacement of proteasome subunits X and Y by LMP7 and LMP2 induced by interferon-gamma for acquirement of the functional diversity responsible for antigen processing. FEBS Lett 343:85-88 (1994).

2. Heink S et al. Tumor cell lines expressing the proteasome subunit isoform LMP7E1 exhibit immunoproteasome deficiency. Cancer Res 66:649-652 (2006).

Sequence Similarity

Belongs to the peptidase T1B family.

Post-translational Modification

Autocleaved. The resulting N-terminal Thr residue of the mature subunit is responsible for the nucleophile proteolytic activity.

Subcellular Location

Nucleus. Cytoplasm.

UNIPROT

SwissProt: P28062 Human

SwissProt: P28063 Mouse

SwissProt: P28064 Rat

Synonyms

ALDD antibody

D6S216 antibody

D6S216E antibody

Large multifunctional peptidase 7 antibody

Large multifunctional protease 7 antibody

LMP 7 antibody

LMP7 antibody

Low molecular mass protein 7 antibody

Low molecular weight protein 7 antibody

Macropain subunit C13 antibody

Expand

Images

  • Western blot analysis of Proteasome 20S LMP7 on different lysates with Rabbit anti-Proteasome 20S LMP7 antibody (<a href="/products/ET7107-36" style="font-weight: bold;text-decoration: underline;">ET7107-36</a>) at 1/50,000 dilution.<br /> <br />Lane 1: Raji cell lysate<br />Lane 2: HL-60 cell lysate<br />Lane 3: U-937 cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /> <br />Predicted band size: 23 kDa<br />Observed band size: 23 kDa<br /> <br />Exposure time: 3 minute; ECL: K1801;<br />4-20% SDS-PAGE gel.<br /> <br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET7107-36" style="font-weight: bold;text-decoration: underline;">ET7107-36</a>) at 1/50,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of Proteasome 20S LMP7 on different lysates with Rabbit anti-Proteasome 20S LMP7 antibody (ET7107-36) at 1/50,000 dilution.

    Lane 1: Raji cell lysate
    Lane 2: HL-60 cell lysate
    Lane 3: U-937 cell lysate

    Lysates/proteins at 20 µg/Lane.

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

    Exposure time: 3 minute; ECL: K1801;
    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET7107-36) at 1/50,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:50,000 dilution was used for 1 hour at room temperature.

  • Immunocytochemistry analysis of U-937 cells labeling Proteasome 20S LMP7 with Rabbit anti-Proteasome 20S LMP7 antibody (<a href="/products/ET7107-36" style="font-weight: bold;text-decoration: underline;">ET7107-36</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Proteasome 20S LMP7 antibody (<a href="/products/ET7107-36" style="font-weight: bold;text-decoration: underline;">ET7107-36</a>) at 1/100 dilution in 1% BSA in PBST 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.<br /><br />Beta tubulin (<a href="/products/HA601187" style="font-weight: bold;text-decoration: underline;">HA601187</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of U-937 cells labeling Proteasome 20S LMP7 with Rabbit anti-Proteasome 20S LMP7 antibody (ET7107-36) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Proteasome 20S LMP7 antibody (ET7107-36) at 1/100 dilution in 1% BSA in PBST 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.

    Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-Proteasome 20S LMP7 antibody (<a href="/products/ET7107-36" style="font-weight: bold;text-decoration: underline;">ET7107-36</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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/ET7107-36" style="font-weight: bold;text-decoration: underline;">ET7107-36</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 human kidney tissue with Rabbit anti-Proteasome 20S LMP7 antibody (ET7107-36) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (ET7107-36) 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-Proteasome 20S LMP7 antibody (<a href="/products/ET7107-36" style="font-weight: bold;text-decoration: underline;">ET7107-36</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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/ET7107-36" style="font-weight: bold;text-decoration: underline;">ET7107-36</a>) at 1/200 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-Proteasome 20S LMP7 antibody (ET7107-36) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (ET7107-36) at 1/200 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 small intestine tissue with Rabbit anti-Proteasome 20S LMP7 antibody (<a href="/products/ET7107-36" style="font-weight: bold;text-decoration: underline;">ET7107-36</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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/ET7107-36" style="font-weight: bold;text-decoration: underline;">ET7107-36</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 small intestine tissue with Rabbit anti-Proteasome 20S LMP7 antibody (ET7107-36) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (ET7107-36) 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-Proteasome 20S LMP7 antibody (<a href="/products/ET7107-36" style="font-weight: bold;text-decoration: underline;">ET7107-36</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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/ET7107-36" style="font-weight: bold;text-decoration: underline;">ET7107-36</a>) at 1/200 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-Proteasome 20S LMP7 antibody (ET7107-36) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (ET7107-36) at 1/200 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 U-937 cells labeling Proteasome 20S LMP7.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/ET7107-36" style="font-weight: bold;text-decoration: underline;">ET7107-36</a>, 1μg/mL) (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 U-937 cells labeling Proteasome 20S LMP7.

    Cells were fixed and permeabilized. Then stained with the primary antibody (ET7107-36, 1μg/mL) (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).

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

Citation