Anti-SAH Antibody [839-6] (A51758)

Immunofluorescence LSCM results of normal liver cells L02 cultured in RPMI 1640 with 10% FBS for 16h followed by stimulation by 0.5 mM methionine for 24h (A1-A4) double stained with MAF00302 at 40 µg/ml and MAF00202 at 40 µg/ml followed by DAPI staining. Hepatocellular carcinoma cell line HepG2 cells were cultured in RPMI 1640 with 10% FBS for 16h followed by stimulation by 0.5 mM methionine for 24h (B1 -B4) and double stained with MAF00302 at 60 µg/ml and MAF00202 at 60 µg/ml followed by DAPI staining. Different views are as follows: DAPI (A1, B1), MAF00302 (A2, B2) MAF00202 (A3, B3); Overlap of all the three fluorescent signals (A4, B4). Expression patterns of SAM and SAH are different between L02 and HepG2 cells after methionine stimulation for 24h. In the case of cells not actively proliferating, both SAM and SAH are seen more in cytoplasm (more in mitochondria areas) than nuclear.

Flow Cytometry of L02 (A1-A3) and HepG2 (B1-B3) cells double stained with MAF00302 at 18 µg/ml and MAF00201 at 4.5 µg/ml. Colour legend: Orange: blank; Blue: Nuclear fixation / permeabilisation buffer was used (eBioscience 00-5523 FoxP3_TF Staining Buffer Set); Red: Intracellular fixation / permeabilisation buffer was used (eBioscience 00-8824). 100% confluent cells (cultured for 48h) were fixed and permeabilised with the intracellular fixation / permeabilisation buffer (A2, B2) or the nuclear fixation / permeabilisation buffer (A3, B3) and then double stained with the antibodies indicated above. Cells were used for analysis with BD FACS Calibur Flow Cytometer. SAM expression is higher in L02 than HepG2 cells. Both SAM and SAH are expressed ubiquitously yet rather dynamically. The level of SAM is higher than that of SAH in both cells.