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COASY Recombinant Rabbit Monoclonal Antibody [JE58-56]

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Specification

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Catalog# HA721128

COASY Recombinant Rabbit Monoclonal Antibody [JE58-56]

Application

  • WB

  • IF-Cell

  • FC

Reactivity

  • Human

Conjugation

  • unconjugated

Overview

Product Name

COASY Recombinant Rabbit Monoclonal Antibody [JE58-56]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human COASY aa 510-564.

Species Reactivity

Human

Validated Applications

WB, IF-Cell, FC

Molecular Weight

Predicted band size: 62 kDa

Positive Control

HepG2 cell lysate, HeLa cell lysate, A431 cell lysate, HepG2.

Conjugation

unconjugated

Clone Number

JE58-56

RRID

AB_3072252

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

  • 1:1,000-1:2,000

  • IF-Cell

  • 1:100

  • FC

  • 1:1,000

Target

Function

Coenzyme A (CoA) functions as a carrier of acetyl and acyl groups in cells and thus plays an important role in numerous synthetic and degradative metabolic pathways in all organisms. In eukaryotes, CoA and its derivatives are also involved in membrane trafficking and signal transduction. This gene encodes the bifunctional protein coenzyme A synthase (CoAsy) which carries out the last two steps in the biosynthesis of CoA from pantothenic acid (vitamin B5). The phosphopantetheine adenylyltransferase domain of this bifunctional protein catalyzes the conversion of 4'-phosphopantetheine into dephospho-coenzyme A (dpCoA) while its dephospho-CoA kinase domain completes the final step by phosphorylating dpCoA to form CoA. Mutations in this gene are associated with neurodegeneration with brain iron accumulation (NBIA). Alternative splicing results in multiple isoforms. Bifunctional enzyme that catalyzes the fourth and fifth sequential steps of CoA biosynthetic pathway. The fourth reaction is catalyzed by the phosphopantetheine adenylyltransferase, coded by the coaD domain; the fifth reaction is catalyzed by the dephospho-CoA kinase, coded by the coaE domain. May act as a point of CoA biosynthesis regulation.

Background References

1. Ferrandon S. et. al. CoA Synthase (COASY) Mediates Radiation Resistance via PI3K Signaling in Rectal Cancer. Cancer Res. 2020 Jan

2. Kharabsheh HA. et. al. CoAsy knockdown in TNBC cell lines resulted in no overt effect on cell proliferation in vitro. Biochem Biophys Res Commun. 2020

Subcellular Location

Mitochondrion matrix, Cytoplasm.

UNIPROT

SwissProt: Q13057 Human

Synonyms

Bifunctional coenzyme A synthase antibody

Bifunctional phosphopantetheine adenylyl transferase/dephospho CoA kinase antibody

CoA synthase antibody

COASY antibody

COASY_HUMAN antibody

Coenzyme A synthase antibody

Dephospho-CoA kinase antibody

Dephospho-CoA pyrophosphorylase antibody

Dephosphocoenzyme A kinase antibody

DPCK antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of COASY on different lysates with Rabbit anti-COASY antibody (<a href="/products/HA721128" style="font-weight: bold;text-decoration: underline;">HA721128</a>) at 1/1,000 dilution.<br /><br />Lane 1: HAP1-parental cell lysate<br />Lane 2: HAP1-COASY KD cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 62 kDa<br />Observed band size: 62 kDa<br /><br />Exposure time: 120 seconds; ECL: K1801;<br /><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/HA721128" style="font-weight: bold;text-decoration: underline;">HA721128</a>) at 1/1,000 dilution was used in <a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a> 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.

    ☑ Knockdown (KD)

    Western blot analysis of COASY on different lysates with Rabbit anti-COASY antibody (HA721128) at 1/1,000 dilution.

    Lane 1: HAP1-parental cell lysate
    Lane 2: HAP1-COASY KD cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 62 kDa
    Observed band size: 62 kDa

    Exposure time: 120 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 (HA721128) at 1/1,000 dilution was used in K1803 at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Western blot analysis of COASY on different lysates with Rabbit anti-COASY antibody (<a href="/products/HA721128" style="font-weight: bold;text-decoration: underline;">HA721128</a>) at 1/2,000 dilution.<br /><br />Lane 1: HepG2 cell lysate<br />Lane 2: HeLa cell lysate<br />Lane 3: A431 cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 62 kDa<br />Observed band size: 62 kDa<br /><br />Exposure time: 25 seconds; ECL: K1801;<br /><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/HA721128" style="font-weight: bold;text-decoration: underline;">HA721128</a>) at 1/2,000 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 COASY on different lysates with Rabbit anti-COASY antibody (HA721128) at 1/2,000 dilution.

    Lane 1: HepG2 cell lysate
    Lane 2: HeLa cell lysate
    Lane 3: A431 cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 62 kDa
    Observed band size: 62 kDa

    Exposure time: 25 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 (HA721128) at 1/2,000 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 HepG2 cells labeling COASY with Rabbit anti-COASY antibody (<a href="/products/HA721128" style="font-weight: bold;text-decoration: underline;">HA721128</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-COASY antibody (<a href="/products/HA721128" style="font-weight: bold;text-decoration: underline;">HA721128</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. Counterstained with Mitotracker. Nuclear DNA was labelled in blue with DAPI.

    Immunocytochemistry analysis of HepG2 cells labeling COASY with Rabbit anti-COASY antibody (HA721128) 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-COASY antibody (HA721128) 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. Counterstained with Mitotracker. Nuclear DNA was labelled in blue with DAPI.

  • Flow cytometric analysis of HepG2 cells labeling COASY.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/HA721128" style="font-weight: bold;text-decoration: underline;">HA721128</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 HepG2 cells labeling COASY.

    Cells were fixed and permeabilized. Then stained with the primary antibody (HA721128, 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).

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