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    glutathione-synthetase-antibody-epr6562-ab124811.pdf

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Signal Transduction Metabolism Amino Acids
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Glutathione Synthetase antibody [EPR6562] (ab124811)

  • Datasheet
  • SDS
Reviews (6) Submit a question References (6)

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Western blot - Anti-Glutathione Synthetase antibody [EPR6562] (ab124811)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glutathione Synthetase antibody [EPR6562] (ab124811)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glutathione Synthetase antibody [EPR6562] (ab124811)
  • Western blot - Anti-Glutathione Synthetase antibody [EPR6562] (ab124811)
  • Anti-Glutathione Synthetase antibody [EPR6562] (ab124811)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR6562] to Glutathione Synthetase
  • Suitable for: WB, IHC-P
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

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Recombinant Human Glutathione Synthetase protein (BSA and azide free) (ab172161)
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Anti-Nrf2 antibody [EP1808Y] - BSA and Azide free (ab180845)

View more associated products

Overview

  • Product name

    Anti-Glutathione Synthetase antibody [EPR6562]
    See all Glutathione Synthetase primary antibodies
  • Description

    Rabbit monoclonal [EPR6562] to Glutathione Synthetase
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-Pmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human Glutathione Synthetase. The exact sequence is proprietary.

  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
  • Dissociation constant (KD)

    KD = 5.70 x 10 -11 M
    Learn more about KD
  • Storage buffer

    pH: 7.20
    Preservative: 0.05% Sodium azide
    Constituents: 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant
  • Concentration information loading...
  • Purity

    Protein A purified
  • Purification notes

    ab124811 is not purified, it is provided in storage buffer.
  • Clonality

    Monoclonal
  • Clone number

    EPR6562
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Metabolism
    • Amino Acids
    • Neuroscience
    • Neurology process
    • Neurogenesis
    • Signal Transduction
    • Metabolism
    • Drug metabolism
    • Cell Biology
    • Other Antibodies
    • Oxidative Stress
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Amino acid metabolism
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Drug metabolism
    • Metabolism
    • Pathways and Processes
    • Redox metabolism
    • Oxidative stress
    • Metabolism
    • Types of disease
    • Cancer

Associated products

  • Alternative Versions

    • Anti-Glutathione Synthetase antibody [EPR6562] - BSA and Azide free (ab248015)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human GSS (Glutathione Synthetase) knockout HEK-293T cell line (ab266342)
  • KO cell lysates

    • Human GSS (Glutathione Synthetase) knockout HEK-293T cell lysate (ab257460)
  • Positive Controls

    • HeLa whole cell lysate (ab29545)
    • SW480 whole cell lysate (ab3957)
  • Recombinant Protein

    • Recombinant Human Glutathione Synthetase protein (BSA and azide free) (ab172161)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab124811 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB (3)
1/10000 - 1/50000. Detects a band of approximately 52 kDa (predicted molecular weight: 52 kDa).
IHC-P (1)
1/8000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

(Heat to 98&degC, allow to cool for 10-20 minutes)

For unpurified use at dilution between 1/50 and 1/100

Notes
WB
1/10000 - 1/50000. Detects a band of approximately 52 kDa (predicted molecular weight: 52 kDa).
IHC-P
1/8000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

(Heat to 98&degC, allow to cool for 10-20 minutes)

For unpurified use at dilution between 1/50 and 1/100

Target

  • Pathway

    Sulfur metabolism; glutathione biosynthesis; glutathione from L-cysteine and L-glutamate: step 2/2.
  • Involvement in disease

    Defects in GSS are the cause of glutathione synthetase deficiency (GSS deficiency) [MIM:266130]; also known as 5-oxoprolinuria or pyroglutamic aciduria. It is a severe form characterized by an increased rate of hemolysis and defective function of the central nervous system.
    Defects in GSS are the cause of glutathione synthetase deficiency of erythrocytes (GLUSYNDE)[MIM:231900]. Glutathione synthetase deficiency of erythrocytes is a mild form causing hemolytic anemia.
  • Sequence similarities

    Belongs to the eukaryotic GSH synthase family.
  • Target information above from: UniProt accession P48637 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 2937 Human
    • Omim: 601002 Human
    • SwissProt: P48637 Human
    • Unigene: 82327 Human
    • Alternative names

      • 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
      • HEL-S-64p antibody
      • HEL-S-88n antibody
      • MGC14098 antibody
      • OTTHUMP00000030711 antibody
      see all

    Images

    • Western blot - Anti-Glutathione Synthetase antibody [EPR6562] (ab124811)
      Western blot - Anti-Glutathione Synthetase antibody [EPR6562] (ab124811)
      All lanes : Anti-Glutathione Synthetase antibody [EPR6562] (ab124811) at 1/10000 dilution (Purified)

      Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
      Lane 2 : 293T (Human embryonic kidney epithelial cell) whole cell lysate
      Lane 3 : HT-1080 (Human fibrosarcoma epithelial cell) whole cell lysate

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 52 kDa

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glutathione Synthetase antibody [EPR6562] (ab124811)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glutathione Synthetase antibody [EPR6562] (ab124811)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast tissue sections labeling Glutathione Synthetase with Purified ab124811 at 1:8000 (0.11 µg/ml). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) . Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control. 

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glutathione Synthetase antibody [EPR6562] (ab124811)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glutathione Synthetase antibody [EPR6562] (ab124811)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human hepatocellular carcinoma tissue sections labeling Glutathione Synthetase with Purified ab124811 at 1:8000 (0.11 µg/ml). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) . Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control. 

    • Western blot - Anti-Glutathione Synthetase antibody [EPR6562] (ab124811)
      Western blot - Anti-Glutathione Synthetase antibody [EPR6562] (ab124811)
      All lanes : Anti-Glutathione Synthetase antibody [EPR6562] (ab124811) at 1/10000 dilution

      Lane 1 : Wild-type HEK-293T cell lysate
      Lane 2 : GSS knockout HEK-293T cell lysate
      Lane 3 : HeLa cell lysate
      Lane 4 : Daudi cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

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



      Lanes 1- 4: Merged signal (red and green). Green - ab124811 observed at 52 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

       ab124811 was shown to react with GSS in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266342 (knockout cell lysate ab257460) was used. Wild-type HEK-293T and GSS knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab124811 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 10000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

    • Anti-Glutathione Synthetase antibody [EPR6562] (ab124811)
      Anti-Glutathione Synthetase antibody [EPR6562] (ab124811)

    Protocols

    • Immunohistochemistry protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (6)

    Publishing research using ab124811? Please let us know so that we can cite the reference in this datasheet.

    ab124811 has been referenced in 6 publications.

    • Tang Z  et al. Deubiquitinase USP35 modulates ferroptosis in lung cancer via targeting ferroportin. Clin Transl Med 11:e390 (2021). PubMed: 33931967
    • Yang Y  et al. RRM2 protects against ferroptosis and is a tumor biomarker for liver cancer. Cancer Cell Int 20:587 (2020). PubMed: 33372599
    • Nishizawa S  et al. Low tumor glutathione level as a sensitivity marker for glutamate-cysteine ligase inhibitors. Oncol Lett 15:8735-8743 (2018). PubMed: 29928324
    • Liu C  et al. Diallyl Trisulfide Protects Motor Neurons from the Neurotoxic Protein TDP-43 via Activating Lysosomal Degradation and the Antioxidant Response. Neurochem Res 43:2304-2312 (2018). PubMed: 30317421
    • Parsanathan R & Jain SK Hydrogen sulfide increases glutathione biosynthesis, and glucose uptake and utilisation in C2C12 mouse myotubes. Free Radic Res 52:288-303 (2018). PubMed: 29378451
    • Wang C  et al. Acetaminophen attenuates glomerulosclerosis in obese Zucker rats via reactive oxygen species/p38MAPK signaling pathways. Free Radic Biol Med 81:47-57 (2015). PubMed: 25614458

    Customer reviews and Q&As

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    1-6 of 6 Abreviews or Q&A

    Western blot abreview for Anti-Glutathione Synthetase antibody [EPR6562]

    Poor
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (Kidney)
    Gel Running Conditions
    Reduced Denaturing (8%)
    Loading amount
    50 µg
    Specification
    Kidney
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Dr. Rami Massri

    Verified customer

    Submitted Feb 24 2021

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Glutathione Synthetase antibody [EPR6562]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (MDA-MB-468 tumor taken out of Balb/C nude mice)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citrate pH 6
    Permeabilization
    No
    Specification
    MDA-MB-468 tumor taken out of Balb/C nude mice
    Blocking step
    Innovex Background Buster as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 20°C
    Fixative
    Formaldehyde
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Jun 22 2018

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Glutathione Synthetase antibody [EPR6562]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (U937 cells)
    Permeabilization
    Yes - 0.1% TritonX-100
    Specification
    U937 cells
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 20°C
    Fixative
    Formaldehyde
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted May 24 2018

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Glutathione Synthetase antibody [EPR6562]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Mouse Cell (B16 cells)
    Permeabilization
    Yes - 0.1% TritonX-100
    Specification
    B16 cells
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 20°C
    Fixative
    Formaldehyde
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted May 24 2018

    Western blot abreview for Anti-Glutathione Synthetase antibody [EPR6562]

    Inconclusive
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (B16 tumor lysate)
    Gel Running Conditions
    Reduced Denaturing (4-20% TG gels)
    Loading amount
    50 µg
    Specification
    B16 tumor lysate
    Blocking step
    Milk as blocking agent for 18 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Apr 10 2018

    Western blot abreview for Anti-Glutathione Synthetase antibody [EPR6562]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (THP1 cells)
    Gel Running Conditions
    Reduced Denaturing (4-20% Tris Glycin)
    Loading amount
    50 µg
    Specification
    THP1 cells
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Feb 21 2018

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