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Glutathione Synthetase Recombinant Rabbit Monoclonal Antibody [JB95-33]

Usd: 385

Specification

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Catalog# ET7107-62

Glutathione Synthetase Recombinant Rabbit Monoclonal Antibody [JB95-33]

Application

  • WB

  • IHC-P

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

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

Overview

Product Name

Glutathione Synthetase Recombinant Rabbit Monoclonal Antibody [JB95-33]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within Human Glutathione Synthetase aa 301-420 / 474.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IHC-P

Target Molecular Weight

Predicted band size: 52 kDa

Positive Control

HeLa cell lysate, Human liver tissue lysate, Mouse kidney tissue lysate, Rat kidney tissue lysate, human kidney tissue, mouse kidney tissue, rat kidney tissue.

Conjugation

unconjugated

Clone Number

JB95-33

RRID

AB_3070771

Product Features

Form

Liquid

Concentration

1 mg/mL.(The concentration of this product may be batch-dependent)

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

Target

Function

GSS (Glutathione synthetase) is a 474 amino acid protein encoded by the gene located at human chromosome 20q11.2. GSS consists of three loops projecting from an antiparallel β-sheet, a parallel β-sheet and a lid of anti-parallel sheets, which provide access to the ATP-binding site. Although Southern blot and gene analysis suggest that GSS may be the only member of a unique family, the crystal structure indicates that GSS belongs to the ATP-GRASP superfamily. GSS is expressed in hemocytes and nucleated cells, including the brain. GSS occurs as a homodimer. There are two steps in the production of Glutathione, begining with γ-GCS and ending with GSS. In an ATP-dependent reaction, GSS produces Glutathione from γ-glutamylcysteine and glycine precursors. Partial hepatectomy, diethyl maleate, buthionine sulfoximine, tert-butylhaydroquinone and thioacetamide increase the ex-pression of GSS, which causes an increase in Glutathione levels. An inherited autosomal recessive disorder, 5-oxoprolinuria (pyroglutamic aciduria), is caused by GSS deficiencies, which leads to central nervous system damage, hemolytic anemia, metabolic acidosis and urinary excretion of 5-oxoproline.

Background References

1. Polekhina G et al. Molecular basis of glutathione synthetase deficiency and a rare gene permutation event. EMBO J 18:3204-3213 (1999).

2. Shi Z-Z et al. Mutations in the glutathione synthetase gene cause 5-oxoprolinuria. Nat Genet 14:361-365 (1996).

Sequence Similarity

Belongs to the eukaryotic GSH synthase family.

Subcellular Location

Cytosol. Extracellular region or secreted.

UNIPROT

SwissProt: P48637 Human

SwissProt: P51855 Mouse

SwissProt: P46413 Rat

Synonyms

epididymis secretory sperm binding protein Li 64p antibody

epididymis secretory sperm binding protein Li 88n antibody

Glutathione synthase antibody

Glutathione synthetase antibody

GSH S antibody

GSH synthetase antibody

GSH-S antibody

GSHB_HUMAN antibody

GSHS antibody

GSS antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of Glutathione Synthetase on different lysates with Rabbit anti-Glutathione Synthetase antibody (<a href="/products/ET7107-62" style="font-weight: bold;text-decoration: underline;">ET7107-62</a>) at 1/1,000 dilution.<br /><br />Lane 1: HAP1-parental cell lysate<br />Lane 2: HAP1-Glutathione Synthetase KD cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 52 kDa<br />Observed band size: 52 kDa<br /><br />Exposure time: 180 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/ET7107-62" style="font-weight: bold;text-decoration: underline;">ET7107-62</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 Glutathione Synthetase on different lysates with Rabbit anti-Glutathione Synthetase antibody (ET7107-62) at 1/1,000 dilution.

    Lane 1: HAP1-parental cell lysate
    Lane 2: HAP1-Glutathione Synthetase KD cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 52 kDa
    Observed band size: 52 kDa

    Exposure time: 180 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 (ET7107-62) 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 Glutathione Synthetase on different lysates with Rabbit anti-Glutathione Synthetase antibody (<a href="/products/ET7107-62" style="font-weight: bold;text-decoration: underline;">ET7107-62</a>) at 1/1,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: Human liver tissue lysate<br />Lane 3: Mouse kidney tissue lysate<br />Lane 4: Rat kidney tissue lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 52 kDa<br />Observed band size: 52 kDa<br /><br />Exposure time: Lane 1-2: 3 minutes; Lane 3-4: 30 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/ET7107-62" style="font-weight: bold;text-decoration: underline;">ET7107-62</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 Glutathione Synthetase on different lysates with Rabbit anti-Glutathione Synthetase antibody (ET7107-62) at 1/1,000 dilution.

    Lane 1: HeLa cell lysate
    Lane 2: Human liver tissue lysate
    Lane 3: Mouse kidney tissue lysate
    Lane 4: Rat kidney tissue lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 52 kDa
    Observed band size: 52 kDa

    Exposure time: Lane 1-2: 3 minutes; Lane 3-4: 30 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 (ET7107-62) 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 kidney tissue with Rabbit anti-Glutathione Synthetase antibody (<a href="/products/ET7107-62" style="font-weight: bold;text-decoration: underline;">ET7107-62</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/ET7107-62" style="font-weight: bold;text-decoration: underline;">ET7107-62</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-Glutathione Synthetase antibody (ET7107-62) 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 (ET7107-62) 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-Glutathione Synthetase antibody (<a href="/products/ET7107-62" style="font-weight: bold;text-decoration: underline;">ET7107-62</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/ET7107-62" style="font-weight: bold;text-decoration: underline;">ET7107-62</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 kidney tissue with Rabbit anti-Glutathione Synthetase antibody (ET7107-62) 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 (ET7107-62) 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-Glutathione Synthetase antibody (<a href="/products/ET7107-62" style="font-weight: bold;text-decoration: underline;">ET7107-62</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/ET7107-62" style="font-weight: bold;text-decoration: underline;">ET7107-62</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 kidney tissue with Rabbit anti-Glutathione Synthetase antibody (ET7107-62) 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 (ET7107-62) 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.

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Citation