Cat. # | Size | Qty. | Price |
---|---|---|---|
48005S | 100 µl |
|
REACTIVITY | H |
SENSITIVITY | Endogenous |
MW (kDa) | 78 |
SOURCE | Rabbit |
Product Information
Application | Dilution |
---|---|
Western Blotting | 1:1000 |
For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
NOTE: Please refer to primary antibody product webpage for recommended antibody dilution.
From sample preparation to detection, the reagents you need for your Western Blot are now in one convenient kit: #12957 Western Blotting Application Solutions Kit
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
Load 20 µl onto SDS-PAGE gel (10 cm x 10 cm).
NOTE: Loading of prestained molecular weight markers (#59329, 10 µl/lane) to verify electrotransfer and biotinylated protein ladder (#7727, 10 µl/lane) to determine molecular weights are recommended.
NOTE: Volumes are for 10 cm x 10 cm (100 cm2) of membrane; for different sized membranes, adjust volumes accordingly.
* Avoid repeated exposure to skin.
posted June 2005
revised June 2020
Protocol Id: 10
Human
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human GCLC protein. Antibodies are purified by peptide affinity chromatography.
Glutamate-cysteine ligase catalytic subunit (GCLC) catalyzes the ligation of glutamate and cysteine, the first and rate-limiting step in glutathione biosynthesis (1). Studies show that deubiquitinases protect cancer cells when glutathione synthesis is suppressed. Upon inhibition of both GCLC and deubiquitinases, cancer cells undergo proteotoxic stress, leading to cell death (2). In addition, deletion of Gclc enhances reactive oxygen species in mouse regulatory T cells (Tregs). Glutathione synthesized in part by Gclc is required for the suppressive function of mouse Tregs (3). Furthermore, in non-small cell lung cancer (NSCLC) cell lines, the transcription factor NRF2 stimulates GCLC expression. When NSCLC cells are starved for cysteine, GCLC, through its non-canonical activity, catalyzes the synthesis of γ-glutamyl-peptide, therefore reducing glutamate levels to protect cells against ferroptosis (1).
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