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PPCS Recombinant Rabbit Monoclonal Antibody [JE63-91]

Usd: 385

Specification

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Catalog# HA722435

PPCS Recombinant Rabbit Monoclonal Antibody [JE63-91]

Application

  • WB

  • IHC-P

  • IF-Tissue

  • FC

  • IP

Reactivity

  • Human

Conjugation

  • unconjugated

Overview

Product Name

PPCS Recombinant Rabbit Monoclonal Antibody [JE63-91]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human PPCS aa 262-311 / 311.

Species Reactivity

Human

Validated Applications

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

Molecular Weight

Predicted band size: 34 kDa

Positive Control

HeLa cell lysate, 293T cell lysate, SH-SY5Y cell lysate, HepG2 cell lysate, A431 cell lysate, human colon tissue, human colon cancer tissue, human kidney tissue, A431.

Conjugation

unconjugated

Clone Number

JE63-91

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

  • 1:1,000

  • IHC-P

  • 1:200-1:1,000

  • IF-Tissue

  • 1:200

  • FC

  • 1:1,000

  • IP

  • 1-2μg/sample

Target

Function

In enzymology, a phosphopantothenate—cysteine ligase also known as phosphopantothenoylcysteine synthetase (PPCS) is an enzyme (EC 6.3.2.5) that catalyzes the chemical reaction which constitutes the second of five steps involved in the conversion of pantothenate to Coenzyme A. The nucleoside triphosphate (NTP) involved in the reaction varies from species to species. Phosphopantothenate—cysteine ligase from the bacterium Escherichia coli uses cytidine triphosphate (CTP) as an energy donor, whilst the human isoform uses adenosine triphosphate (ATP).

Background References

1. Evans JC et al. Targeting Mycobacterium tuberculosis CoaBC through Chemical Inhibition of 4\'-Phosphopantothenoyl-l-cysteine Synthetase (CoaB) Activity. ACS Infect Dis. 2021 Jun

2. Lok A et al. Novel phosphopantothenoylcysteine synthetase (PPCS) mutations with prominent neuromuscular features: Expanding the phenotypical spectrum of PPCS-related disorders. Am J Med Genet A. 2022 Sep

Subcellular Location

Cytoplasm, cytosol, nucleus.

UNIPROT

SwissProt: Q9HAB8 Human

Synonyms

COAB antibody

FLJ11838 antibody

MGC117357 antibody

MGC138220 antibody

OTTHUMP00000008433 antibody

Phosphopantothenate--cysteine ligase antibody

Phosphopantothenoylcysteine synthetase antibody

PPC synthetase antibody

Ppcs antibody

PPCS_HUMAN antibody

Expand

Images

  • Western blot analysis of PPCS on different lysates with Rabbit anti-PPCS antibody (<a href="/products/HA722435" style="font-weight: bold;text-decoration: underline;">HA722435</a>) at 1/1,000 dilution.<br /><br />Lane 1: HeLa cell lysate (20 µg/Lane)<br />Lane 2: 293T cell lysate (20 µg/Lane)<br />Lane 3: SH-SY5Y cell lysate (20 µg/Lane)<br />Lane 4: HepG2 cell lysate (20 µg/Lane)<br />Lane 5: A431 cell lysate (20 µg/Lane)<br /><br />Predicted band size: 34 kDa<br />Observed band size: 34 kDa<br /><br />Exposure time: 10 seconds; 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/HA722435" style="font-weight: bold;text-decoration: underline;">HA722435</a>) at 1/1,000 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 PPCS on different lysates with Rabbit anti-PPCS antibody (HA722435) at 1/1,000 dilution.

    Lane 1: HeLa cell lysate (20 µg/Lane)
    Lane 2: 293T cell lysate (20 µg/Lane)
    Lane 3: SH-SY5Y cell lysate (20 µg/Lane)
    Lane 4: HepG2 cell lysate (20 µg/Lane)
    Lane 5: A431 cell lysate (20 µg/Lane)

    Predicted band size: 34 kDa
    Observed band size: 34 kDa

    Exposure time: 10 seconds; 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 (HA722435) at 1/1,000 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.

  • Immunohistochemical analysis of paraffin-embedded human colon tissue with Rabbit anti-PPCS antibody (<a href="/products/HA722435" style="font-weight: bold;text-decoration: underline;">HA722435</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/HA722435" style="font-weight: bold;text-decoration: underline;">HA722435</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 colon tissue with Rabbit anti-PPCS antibody (HA722435) 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 (HA722435) 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 colon cancer tissue with Rabbit anti-PPCS antibody (<a href="/products/HA722435" style="font-weight: bold;text-decoration: underline;">HA722435</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/HA722435" style="font-weight: bold;text-decoration: underline;">HA722435</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 colon cancer tissue with Rabbit anti-PPCS antibody (HA722435) 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 (HA722435) 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 kidney tissue with Rabbit anti-PPCS antibody (<a href="/products/HA722435" style="font-weight: bold;text-decoration: underline;">HA722435</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/HA722435" style="font-weight: bold;text-decoration: underline;">HA722435</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-PPCS antibody (HA722435) 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 (HA722435) 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 A431 cells labeling PPCS.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/HA722435" style="font-weight: bold;text-decoration: underline;">HA722435</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 A431 cells labeling PPCS.

    Cells were fixed and permeabilized. Then stained with the primary antibody (HA722435, 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).

  • PPCS was immunoprecipitated from 0.2 mg 293T cell lysate with <a href="/products/HA722435" style="font-weight: bold;text-decoration: underline;">HA722435</a> at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using <a href="/products/HA722435" style="font-weight: bold;text-decoration: underline;">HA722435</a> at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.<br /><br />Lane 1: 293T cell lysate (input)<br />Lane 2: <a href="/products/HA722435" style="font-weight: bold;text-decoration: underline;">HA722435</a> IP in 293T cell lysate<br />Lane 3: Rabbit IgG instead of <a href="/products/HA722435" style="font-weight: bold;text-decoration: underline;">HA722435</a> in 293T cell lysate<br /><br />Blocking/Dilution buffer: 5% NFDM/TBST<br />Exposure time: 20 seconds; ECL: <a href="/products/K1801" style="font-weight: bold;text-decoration: underline;">K1801</a>

    PPCS was immunoprecipitated from 0.2 mg 293T cell lysate with HA722435 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA722435 at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.

    Lane 1: 293T cell lysate (input)
    Lane 2: HA722435 IP in 293T cell lysate
    Lane 3: Rabbit IgG instead of HA722435 in 293T cell lysate

    Blocking/Dilution buffer: 5% NFDM/TBST
    Exposure time: 20 seconds; ECL: K1801

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