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CLPX Recombinant Rabbit Monoclonal Antibody [JE62-96]

Usd: 385

Specification

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Catalog# HA721016

CLPX Recombinant Rabbit Monoclonal Antibody [JE62-96]

Application

  • WB

  • IF-Cell

  • IHC-P

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

Overview

Product Name

CLPX Recombinant Rabbit Monoclonal Antibody [JE62-96]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within human CLPX aa 584-633/633.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, IHC-P

Molecular Weight

Predicted band size: 69 kDa

Positive Control

Raji cell lysate, Hela cell lysate, HepG2 cell lysate, NIH/3T3 cell lysate, Rat heart tissue lysate, Mouse heart tissue lysate, Mouse liver tissue lysate, Rat liver tissue lysate, human testis tissue, mouse testis tissue, rat testis tissue, NCI-H441.

Conjugation

unconjugated

Clone Number

JE62-96

RRID

AB_3072140

Product Features

Form

Liquid

Concentration

Storage Instructions

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

  • IF-Cell

  • 1:50

  • IHC-P

  • 1:200-1:1,000

Target

Function

ATP-dependent Clp protease ATP-binding subunit clpX-like, mitochondrial is an enzyme that in humans is encoded by the CLPX gene. This protein is a member of the family of AAA Proteins (AAA+ ATPase) and is to form the protein complex of Clp protease. ClpX is an ATP-dependent chaperone that can recognize protein substrates by binding to protein degradation tags. These tags can be short unstructured peptide sequences (e.g., ssrA-tag in E coli). As an essential component of ClpP protease complex, ClpX recruits degradable substrates and unfolds their tertiary structure, which requires energy provided by ATP hydrolysis. Subsequently, these ClpX Chaperones transfer protein substrates into the proteolytic chamber formed by ClpP tetradecamer. In mammals, ClpXP protease is a pivotal contributor to mitochondrial protein quality control. A compromised ClpXP function usually leads to the accumulation of damaged proteins and mitochondrial dysfunctions, which believes to be potential causes for neurodegenerative diseases and aging.

Background References

1. Kang ZH. et. al. Progress and prospect of single-molecular ClpX ATPase researching system-a mini-review. Gene. 2021 Mar

2. Kester JC. et. al. ClpX Is Essential and Activated by Single-Strand DNA Binding Protein in Mycobacteria. J Bacteriol. 2021 Jan

Subcellular Location

Mitochondrion, mitochondrion nucleoid.

UNIPROT

SwissProt: O76031 Human

SwissProt: Q9JHS4 Mouse

SwissProt: Q5U2U0 Rat

Synonyms

ATP dependent Clp protease ATP binding subunit clpX like, mitochondrial antibody

ATP-dependent Clp protease ATP-binding subunit clpX-like, mitochondrial antibody

AU014732 antibody

Caseinolytic mitochondrial matrix peptidase chaperone subunit antibody

Caseinolytic peptidase X (E.coli) antibody

Caseinolytic protease X antibody

clpX antibody

ClpX caseinolytic peptidase X homolog antibody

ClpX caseinolytic protease X homolog antibody

CLPX_HUMAN antibody

Expand

Images

  • Western blot analysis of CLPX on different lysates with Rabbit anti-CLPX antibody (<a href="/products/HA721016" style="font-weight: bold;text-decoration: underline;">HA721016</a>) at 1/1,000 dilution.<br /><br />Lane 1: Raji cell lysate<br />Lane 2: Hela cell lysate<br />Lane 3: HepG2 cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 69 kDa<br />Observed band size: 69 kDa<br /><br />Exposure time: 2 minutes;<br /><br />10% 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/HA721016" style="font-weight: bold;text-decoration: underline;">HA721016</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:200,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of CLPX on different lysates with Rabbit anti-CLPX antibody (HA721016) at 1/1,000 dilution.

    Lane 1: Raji cell lysate
    Lane 2: Hela cell lysate
    Lane 3: HepG2 cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 69 kDa
    Observed band size: 69 kDa

    Exposure time: 2 minutes;

    10% 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 (HA721016) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.

  • Western blot analysis of CLPX on different lysates with Rabbit anti-CLPX antibody (<a href="/products/HA721016" style="font-weight: bold;text-decoration: underline;">HA721016</a>) at 1/1,000 dilution.<br /><br />Lane 1: NIH/3T3 cell lysate, 10 µg/Lane<br />Lane 2: Rat heart tissue lysate, 20 µg/Lane<br />Lane 3: Mouse heart tissue lysate, 20 µg/Lane<br />Lane 4: Mouse liver tissue lysate, 20 µg/Lane<br />Lane 5: Rat liver tissue lysate, 20 µg/Lane<br /><br />Predicted band size: 69 kDa<br />Observed band size: 69 kDa<br /><br />Exposure time: 2 minutes;<br /><br />8% 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/HA721016" style="font-weight: bold;text-decoration: underline;">HA721016</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:200,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of CLPX on different lysates with Rabbit anti-CLPX antibody (HA721016) at 1/1,000 dilution.

    Lane 1: NIH/3T3 cell lysate, 10 µg/Lane
    Lane 2: Rat heart tissue lysate, 20 µg/Lane
    Lane 3: Mouse heart tissue lysate, 20 µg/Lane
    Lane 4: Mouse liver tissue lysate, 20 µg/Lane
    Lane 5: Rat liver tissue lysate, 20 µg/Lane

    Predicted band size: 69 kDa
    Observed band size: 69 kDa

    Exposure time: 2 minutes;

    8% 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 (HA721016) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.

  • Immunohistochemical analysis of paraffin-embedded human testis tissue with Rabbit anti-CLPX antibody (<a href="/products/HA721016" style="font-weight: bold;text-decoration: underline;">HA721016</a>) at 1/200 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/HA721016" style="font-weight: bold;text-decoration: underline;">HA721016</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 human testis tissue with Rabbit anti-CLPX antibody (HA721016) at 1/200 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 (HA721016) 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 testis tissue with Rabbit anti-CLPX antibody (<a href="/products/HA721016" style="font-weight: bold;text-decoration: underline;">HA721016</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/HA721016" style="font-weight: bold;text-decoration: underline;">HA721016</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 testis tissue with Rabbit anti-CLPX antibody (HA721016) 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 (HA721016) 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 testis tissue with Rabbit anti-CLPX antibody (<a href="/products/HA721016" style="font-weight: bold;text-decoration: underline;">HA721016</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/HA721016" style="font-weight: bold;text-decoration: underline;">HA721016</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 testis tissue with Rabbit anti-CLPX antibody (HA721016) 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 (HA721016) 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.

  • Immunocytochemistry analysis of NCI-H441 cells labeling CLPX with Rabbit anti-CLPX antibody (<a href="/products/HA721016" style="font-weight: bold;text-decoration: underline;">HA721016</a>) at 1/50 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-CLPX antibody (<a href="/products/HA721016" style="font-weight: bold;text-decoration: underline;">HA721016</a>) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (Alexa Fluor&reg; 488) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/200 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 647, <a href="/products/HA1127" style="font-weight: bold;text-decoration: underline;">HA1127</a>) were used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of NCI-H441 cells labeling CLPX with Rabbit anti-CLPX antibody (HA721016) at 1/50 dilution.

    Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-CLPX antibody (HA721016) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (M1305-2, red) was stained at 1/200 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 647, HA1127) were used as the secondary antibody at 1/1,000 dilution.

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