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GCLC Recombinant Rabbit Monoclonal Antibody [JA08-03]

Usd: 385

Specification

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Catalog# ET1704-38

GCLC Recombinant Rabbit Monoclonal Antibody [JA08-03]

Application

  • WB

  • IHC-P

  • FC

Reactivity

  • Human

  • Rat

Conjugation

  • unconjugated

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

Overview

Product Name

GCLC Recombinant Rabbit Monoclonal Antibody [JA08-03]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human GCLC aa 588-637 / 637.

Species Reactivity

Human, Rat

Validated Applications

WB, IHC-P, FC

Molecular Weight

Predicted band size: 73 kDa

Positive Control

A431 cell lysate, A549 cell lysate, Rat kidney tissue lysate, human fetal kidney tissue, human fetal liver tissue, rat kidney tissue, Jurkat.

Conjugation

unconjugated

Clone Number

JA08-03

RRID

AB_3070487

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

  • 1:500-1:1,000

  • IHC-P

  • 1:50-1:200

  • FC

  • 1:50-1:100

Target

Function

The GCLC gene consists of 16 exons and encodes the 636 amino acid protein g-GCSc (g-glutamylcysteine synthetase heavy subunit), also designated g-L-glutamate-L-cysteine ligase catalytic subunit (GLCLC). g-GCSc is expressed in hemocytes, brain, liver and kidney. g-GCSc associates with a regulatory or modifier subunit, g-GCSm (g-glutamylcysteine synthetase light subunit), to form a heterodimer, g-GCS. g-GCS is the first enzyme involved and the rate determining step in glutathione biosynthesis. Oxidants, cadium and methyl mercury upregulate the transcription of g-GCS. H2O2 regulation depends on the Yap1 protein and the presence of glutamate, glutamine and lysine. Cadium regulates transcription through proteins Met-4, Met-31 and Met-32. Cbf1, a DNA binding protein, inhibits transcription of g-GCS. Chemopreventive compounds cause increased levels of g-GCSc in kidney tissues, which may protect against chemically induced carcinogenesis. A His370Leu amino acid change in g-GCSc causes deficiencies in activity which are responsible for hemolytic anemia and low red blood cell glutathione levels.

Background References

1. Sun J. et. al. High GCLC level in tumor tissues is associated with poor prognosis of hepatocellular carcinoma after curative resection. J Cancer. 2019 Jun

2. Jia X. et. al. Down-regulation of GCLC is involved in microcystin-LR-induced malignant transformation of human liver cells. Toxicology. 2019 Jun

Sequence Similarity

Belongs to the glutamate--cysteine ligase type 3 family.

Subcellular Location

Cytoplasm.

UNIPROT

SwissProt: P48506 Human

SwissProt: P19468 Rat

Synonyms

EC 6.3.2.2 antibody

Gamma ECS antibody

Gamma glutamylcysteine synthetase antibody

Gamma-ECS antibod

Gamma-glutamylcysteine synthetase antibody

GCL antibody

Gclc antibody

GCS antibody

GCS heavy chain antibody

GLCL antibody

Expand

Images

  • Western blot analysis of GCLC on different lysates with Rabbit anti-GCLC antibody (<a href="/products/ET1704-38" style="font-weight: bold;text-decoration: underline;">ET1704-38</a>) at 1/1,000 dilution.<br /><br />Lane 1: A431 cell lysate (20 µg/Lane)<br />Lane 2: A549 cell lysate (20 µg/Lane)<br />Lane 3: Rat kidney tissue lysate (40 µg/Lane)<br /><br />Predicted band size: 73 kDa<br />Observed band size: 73 kDa<br /><br />Exposure time: 1 minute; 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/ET1704-38" style="font-weight: bold;text-decoration: underline;">ET1704-38</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 GCLC on different lysates with Rabbit anti-GCLC antibody (ET1704-38) at 1/1,000 dilution.

    Lane 1: A431 cell lysate (20 µg/Lane)
    Lane 2: A549 cell lysate (20 µg/Lane)
    Lane 3: Rat kidney tissue lysate (40 µg/Lane)

    Predicted band size: 73 kDa
    Observed band size: 73 kDa

    Exposure time: 1 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 (ET1704-38) 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.

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of GCLC on different lysates with Rabbit anti-GCLC antibody (<a href="/products/ET1704-38" style="font-weight: bold;text-decoration: underline;">ET1704-38</a>) at 1/5,000 dilution.<br /><br />Lane 1: A549-si NT cell lysate<br />Lane 2: A549-si GCLC cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 73 kDa<br />Observed band size: 73 kDa<br /><br />Exposure time: 2 minutes; 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/ET1704-38" style="font-weight: bold;text-decoration: underline;">ET1704-38</a>) at 1/5,000 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.

    ☑ Knockdown (KD)

    Western blot analysis of GCLC on different lysates with Rabbit anti-GCLC antibody (ET1704-38) at 1/5,000 dilution.

    Lane 1: A549-si NT cell lysate
    Lane 2: A549-si GCLC cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 73 kDa
    Observed band size: 73 kDa

    Exposure time: 2 minutes; 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 (ET1704-38) at 1/5,000 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 fetal kidney tissue with Rabbit anti-GCLC antibody (<a href="/products/ET1704-38" style="font-weight: bold;text-decoration: underline;">ET1704-38</a>) at 1/50 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/ET1704-38" style="font-weight: bold;text-decoration: underline;">ET1704-38</a>) at 1/50 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 fetal kidney tissue with Rabbit anti-GCLC antibody (ET1704-38) at 1/50 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 (ET1704-38) at 1/50 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 fetal liver tissue with Rabbit anti-GCLC antibody (<a href="/products/ET1704-38" style="font-weight: bold;text-decoration: underline;">ET1704-38</a>) at 1/50 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/ET1704-38" style="font-weight: bold;text-decoration: underline;">ET1704-38</a>) at 1/50 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 fetal liver tissue with Rabbit anti-GCLC antibody (ET1704-38) at 1/50 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 (ET1704-38) at 1/50 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-GCLC antibody (<a href="/products/ET1704-38" style="font-weight: bold;text-decoration: underline;">ET1704-38</a>) at 1/50 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/ET1704-38" style="font-weight: bold;text-decoration: underline;">ET1704-38</a>) at 1/50 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-GCLC antibody (ET1704-38) at 1/50 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 (ET1704-38) at 1/50 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 GCLC was done on Jurkat cells. The cells were fixed, permeabilized and stained with the primary antibody (<a href="/products/ET1704-38" style="font-weight: bold;text-decoration: underline;">ET1704-38</a>, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor&reg;488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of GCLC was done on Jurkat cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1704-38, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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