AB5010

Anti-Glutathione Antibody, detects both GSH & GSSG

Chemicon^®, from rabbit

• UNSPSC Code: 12352203
• NACRES: NA.41
• eCl@ss: 32160702
• Clone: polyclonal
• Species reactivity: eukaryotes
• Application: IHC
• Citations: 13

Properties

• biological source: rabbit
• Quality Segment: 100
• antibody form: purified immunoglobulin
• antibody product type: primary antibodies
• clone: polyclonal
• species reactivity: eukaryotes
• manufacturer/tradename: Chemicon^®
• technique(s): immunohistochemistry: suitable
• shipped in: wet ice
• target post-translational modification: unmodified
• Gene Information: human ... GSS(2937)

Description

Immunogen

Epitope: detects both GSH & GSSG

Glutathione-glutaraldehyde

Application

This Anti-Glutathione Antibody, detects both GSH & GSSG is validated for use in IH for the detection of Glutathione.

Immunohistochemistry using silver-intensified immunogold or enhanced fluorescence methods. Treatment with autofluorescent reducers is typically necessary when using glutaraldehyde fixatives. Samples should be fixed with 0.2% - 2.5% glutaraldehyde for optimum detection. See notes technical notes. This antibody has also been used and found to work with a zero-low glutaraldehyde / high paraformaldehyde fixation (4% paraformaldehyde in 0.1M phosphate buffer / 3% sucrose fixative ). The minimum glutaraldehyde concentration for AB5010 is 0.02%. Performance is good with frozen sections, Vibratome sections and tissue culture formats, when penetrating reagents such as 0.3% Triton X-100 are used (only in blocking buffer, antibody dilution buffer should be in simple TBS or PBS only).

Western blot (Mawatari and Murakami, 2004).

Optimal working dilutions must be determined by the end user.

Note: Glutathione is not osmium tolerant and therefore should not be used for post-embedding electron micoscope immunohistochemistry.



All procedures may be carried out at room temperature. Though the exact dilutions for all applications can not be predicated, it is unlikely that deviations from the calibrated concentration will be needed. The 100X dilution is calibrated to optimize the detection of antigen levels over a 2 log unit range. Greater or lesser dilutions may be employed if unusually high (>20 mM) or low (<0.2 mM) intracellular hapten levels are anticipated or novel specimen conditions exist, such as low glutaraldehyde levels or unusual embedding protocols. Greater dilutions will likely result in decreased maximum signal strength whereas more concentrated solutions will compress the dynamic range of detection.



TECHNICAL NOTES

The AB5010 is optimized for use with immunogold or fluorescence detection. Visualization with enzyme-linked methods can be used but they typically compress the dynamic range of detection. Use of high gain methods such as tyramides will require the user to perform independent calibrations. Use with frozen sections is possible but will not yield optimal images as IgGs will penetrate aldehyde cross-linked tissue poorly and most amino acids are present at such high levels that prozone effect occurs. Use in whole mount or vibratome sections is not recommended for similar reasons.

Research Category
Neuroscience

Research Sub Category
Oxidative Stress

Biochem/physiol Actions

Glutathione. The antibody has been calibrated against a spectrum of antigens to assure hapten selectivity and proper affinity. No measurable glutaraldehyde-fixed tissue cross-reactivity (<1:1000) against L-alanine, g-aminobutyrate, 1-amino-4-guanidobutane (AGB), D/L-arganine, D/L-aspartate, L-citrulline, L-cysteine, D/L-glutamate, D/L-glutamine, glycine, L-lysine, L-ornithine, L-serine, taurine, L-threonine, L-tryptophan, L-tyrosine. Does not discriminate between free GSH and GSSG.

Packaging

sufficient for 2000 assays

Physical form

Format: Purified

IgG fraction in sterile 0.1M phosphate buffer. No preservative

Preparation Note

Maintain stock at 2-8°C in undiluted aliquots for up to 6 months. This stock is extremely stable under normal use and routine storage at 2-8°C. Do not refreeze this stock.

Legal Information

CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.