Flow cytometric analysis of Glutamate-ammonia ligase expression in HeLa cells using Glutamate-ammonia ligase antibody (Cat#63474, 1:2,000). Green, isotype control; red, Glutamate-ammonia ligase.
Immunocytochemical staining of HeLa cells with Glutamate-ammonia ligase antibody (Cat#63474, 1:1,000). Nuclei were stained blue with DAPI; Glutamate-ammonia ligase 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.
Western blotting analysis using anti-Glutamate-ammonia ligase antibody (Cat#63474). Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with anti-Glutamate-ammonia ligase antibody (Cat#63474, 1:5,000) and HRP-conjugated goat anti-rabbit secondary antibody (Cat#201, 1:20,000) respectively. Image was developed using FeQ™ ECL Substrate Kit (Cat#226).
Western blotting analysis using anti-Glutamate-ammonia ligase antibody (Cat#63474). Glutamate-ammonia ligase expression in wild type (WT) and Glutamate-ammonia ligase shRNA knockdown (KD) HeLa cells with 30 μg of total cell lysates. β-Tubulin serves as a loading control. The blot was incubated with anti-Glutamate-ammonia ligase antibody (Cat#63474, 1:5,000) and HRP-conjugated goat anti-rabbit secondary antibody (Cat#201, 1:20,000) respectively. Image was developed using FeQ™ ECL Substrate Kit (Cat#226).
Validation of Glutamine Synthetase knockdown using flow cytometry. Wild-type(WT, Blue) and knockdown(KD, Green) HeLa cells were stained with anti-Glutamine Synthetase antibody (Cat#63474, 1:2,000) and analyzed using CytoFLEX.
Immunocytochemical staining of HeLa cells using anti-Glutamate-ammonia ligase antibody (Cat#63474, 1:1,000), Top panel: wild-type (WT); Bottom panal: Glutamate-ammonia ligase shRNA knockdown (KD). Nuclei were stained blue with DAPI; Glutamate-ammonia ligase was stained magenta with Alexa Fluor® 647. Scale bar, 20 μm. Permeabilization: Triton.
Immunohistochemistry was performed on paraffin-embedded human hepatocarcinoma using anti-Glutamate-ammonia ligase antibody (Cat#63474, 1:200). Antigen retrieval was done in sodium citrate buffer (pH 6.0). DAB was used for detection, with hematoxylin counterstaining. Images were acquired using a Nikon Ci-L Plus microscope (40× objective). Scale bar: 25 μm.