Overview

  • Product name

    Anti-Glutathione Synthetase antibody
    See all Glutathione Synthetase primary antibodies
  • Description

    Rabbit polyclonal to Glutathione Synthetase
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, ELISAmore details
  • Species reactivity

    Reacts with: Mouse, Human
  • Immunogen

    KLH conjugated synthetic peptide selected from the C terminal region of Human Glutathione Synthetase.

  • Positive control

    • Mouse liver tissue lysate

Properties

Applications

Our Abpromise guarantee covers the use of ab91591 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 1/100 - 1/500. Predicted molecular weight: 52 kDa.
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ELISA 1/1000.

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.
  • Information by UniProt
  • Database links

  • 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

  • Anti-Glutathione Synthetase antibody (ab91591) at 1/100 dilution + Mouse liver tissue lysate at 35 µg

    Predicted band size: 52 kDa

  • IHC image of ab91591 staining in Human normal liver formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab91591, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

References

This product has been referenced in:

  • Booty LM  et al. Selective Disruption of Mitochondrial Thiol Redox State in Cells and In Vivo. Cell Chem Biol 26:449-461.e8 (2019). Read more (PubMed: 30713096) »
  • Lv W  et al. Schwann Cell Plasticity is Regulated by a Weakened Intrinsic Antioxidant Defense System in Acute Peripheral Nerve Injury. Neuroscience 382:1-13 (2018). Read more (PubMed: 29684504) »
See all 5 Publications for this product

Customer reviews and Q&As

1-4 of 4 Abreviews or Q&A

Application
Western blot
Sample
Human Cell lysate - whole cell (U937 and MDA-MB-231 cells)
Gel Running Conditions
Reduced Denaturing (4-20% Tris Glycin gel, semi dry transfer)
Loading amount
50 µg
Specification
U937 and MDA-MB-231 cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Jun 18 2018

Application
Western blot
Sample
Mouse Cell lysate - whole cell (B16 and EMT6 cells)
Gel Running Conditions
Reduced Denaturing (4-20% Tris Glycin gel, semi dry transfer)
Loading amount
50 µg
Specification
B16 and EMT6 cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Jun 18 2018

Answer

Thank you for contacting us.

Please find the ELISA Protocol below.

Note: The precise conditions should be optimized for a particular assay.

Solution Preparation

Coating Solution:

Antigen is diluted in coating solution for immobilization onto the microplate. Commonly used coating solutions are: 50 mM sodium carbonate, pH 9.6; 20 mM Tris-HCL, pH 8.5; or 10 mM PBS, pH 7.2. An antigen concentration of 1-10 ug/mL is usually sufficient.

Blocking Solution:

Commonly used blocking agents are: BSA, nonfat dry milk, casein, gelatin, etc. Different assay systems may require different blocking agents.

Primary/Secondary Antibody Solution:

Primary/secondary antibody should be diluted in 1x Blocking solution to prevent non-specific binding. It is recommended to diluteprimary antibodies to between 1:100 and 1:1000. Follow the manufacturer’s advice for secondary antibodies.

Wash Solution:

Typically 0.1 M phosphate-buffered saline or Tris-buffered saline (pH 7.4) with a detergent such as Tween 20 (0.02%-0.05% v/v).

Protocol:

Dilute the antigen to 1-2 ug/mL in coating solution
Add 100 uL of diluted antigen to appropriate wells. Incubate 2 hours at room temperature or 4 degrees Celsius overnight.
Empty plate and tap out residual liquid.
Wash twice with 300 uL Wash solution.
Add 300 uL Blocking solution to each well. Incubate 1 hour.
Empty plate and tap out residual liquid.
Wash twice with 300 uL Wash solution.
Add 100 uL diluted primary antibody to each well. Incubate 1 hour at 37 degrees Celsius or 3 hours at room temperature.
Empty plate, tap out residual liquid.
Fill each well with Wash solution. Invert plate to empty, tap out residual liquid. Repeat 3 times.
Add 100 uL diluted secondary antibody to each well. Incubate 1 hour at room temperature.
Empty plate, tap out residual liquid and wash as described in step 10.
Give final 5 minutes soak with Wash solution. Tap residual liquid from plate. This washing step is critical to reduce signal background.
Fill each well with Wash solution. Invert plate to empty, tap out residual liquid. Repeat 5 times.
Dispense 100 uL of substrate (pNPP) into each well. Develop the color for 30 minutes and immediately read plate with plate reader at 405-410 nm.



I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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Read More
Application
Western blot
Sample
Mouse Tissue lysate - whole (Liver)
Loading amount
20 µg
Specification
Liver
Gel Running Conditions
Non-reduced Non-Denaturing (Native) (12)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C

Abcam user community

Verified customer

Submitted Aug 23 2011

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