Technical Data
G2002-01
Gamma Glutamylcysteine Synthetase (GCS, Glutamate-Cysteine Ligase)
Description:
Glutathione (GSH) plays an important role in detoxification of oxidants and toxins from the cells. In most cells GSH is synthesized de novo in two steps catalysed by Gamma Glutamylcysteine Synthetase (GCS) / Glutamate-cystein Ligase and GSH synthetase. GCS catalyzes the first and rate limiting step and is important in GSH homeostasis. It is composed of two subunits: heavy and light. These subunits are coded by different genes which are controlled by different mechanisms. The heavy subunit carries the catalytic activity and can be inhibited through feedback mechanism by GSH. Binding of light subunit to heavy subunit reduces the Michaelis-Menten constant for glutamate to a physiological concentration of 1.2mM and increases the inhibitory constant for GSH.

Applications:
Suitable for use in Western Blot and Immunoprecipitation. Other applications not tested.

Recommended Dilutions:
Western Blot: 2.5-5ug/ml for 2 hours at RT.
Immunoprecipitation: 10ug/mg protein lysate.
Optimal dilutions to be determined by the researcher.

Positive Control:
Jurkat, MAD109, PC12 cells

Storage and Stability:
May be stored at 4C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20C. Aliquots are stable for at least 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
TypeIsotypeCloneGrade
PabIgGAffinity Purified
SizeStorageShippingSourceHost
500ul-20CBlue IceRatRabbit
Concentration:
~1mg/ml
Immunogen:
Synthetic peptide corresponding to aa295-313 of rat Gamma Glutamylcysteine Synthetase. Cellular Localization: Cytoplasmic.
Purity:
Purified by Protein A affinity chromatography.
Form
Supplied as a liquid in 10mM PBS, pH 7.4, 0.2% BSA, 0.09% sodium azide.
Specificity:
Recognizes the heavy subunit of rat Gamma Glutamylcysteine Synthetase at ~73kD. Species Crossreactivity: human, mouse, monkey and bovine (weak).
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
1. Liu R M, et al. (1998) Am J Physiol: Lung Cell Molec Physiol, 275:L861-869.
2. Choi J, et al. (2000) J Biol Chem, 275:3693-3698.
3. Levonen A-L, et al. (2001) Arterioscler Thromb Vasc Biol, 21:1846-1851.
4. Moellering D, et al. (1999) FEBS Letts 448:292-296.