Background

Glutamate-cysteine ligase, also known as gamma-glutamylcysteine synthetase is the first rate-limiting enzyme of glutathione synthesis. The enzyme consists of two subunits, a heavy catalytic subunit and a light regulatory subunit. This locus encodes the catalytic subunit, while the regulatory subunit is derived from a different gene located on chromosome 1p22-p21. Mutations at this locus have been associated with hemolytic anemia due to deficiency of gamma-glutamylcysteine synthetase and susceptibility to myocardial infarction.

Images

	Western blotting analysis using anti-GCLC antibody (Cat#63281). Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with anti-GCLC antibody (Cat#63281, 1:5,000) and HRP-conjugated goat anti-rabbit secondary antibody (Cat#201, 1:20,000) respectively. Image was developed using NaQ™ ECL Substrate Kit (Cat#716) GCLC, glutamate-cysteine ligase catalytic subunit.
	Western blotting analysis using anti-GCLC antibody (Cat#63281). GCLC expression in wild type (WT) and GCLC shRNA knockdown (KD) HeLa cells with 20 μg of total cell lysates. Hsp90 α serves as a loading control. The blot was incubated with anti-GCLC antibody (Cat#63281, 1:5,000) and HRP-conjugated goat anti-rabbit secondary antibody (Cat#201, 1:20,000) respectively. Image was developed using NaQ™ ECL Substrate Kit (Cat#716).
	Flow cytometric analysis of GCLC expression in HepG2 cells using anti-GCLC antibody (Cat#63281, 1:2,000). Green, isotype control; red,GCLC.
	Immunocytochemical staining of HepG2 cells with anti-GCLC antibody (Cat#63281, 1:1,000). Nuclei were stained blue with DAPI; GCLC was stained magenta with Alexa Fluor® 647. Images were taken using Leica stellaris 5. Protein abundance based on laser Intensity and smart gain: Medium. Scale bar: 20 μm.

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