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. Author manuscript; available in PMC: 2012 Nov 1.
Published in final edited form as: J Neurochem. 2011 Oct 11;119(4):749–759. doi: 10.1111/j.1471-4159.2011.07483.x

Figure 4. TLC-immunooverlay of SGPG isolated from SV-HCECs after EGFP, EGFP-GlcATp, EGFP-GlcATs and EGFP-GlcATp + Ts transfection.

Figure 4

The transfected cells were incubated for 48 h. Lipids were extracted using solvent mixtures and SGPG was purified into a fraction using DEAE-Sephadex A25 column. The fraction was dissolved in a defined volume of solvent (chloroform:methanol:water 12:7:1, v/v) according to protein concentration, and a portion of the solution equivalent to an equal amount of protein was applied on an aluminum-backed HPLTC along with a reference standards. The plate was developed in chloroform;methanol:0.25% CaCl2 (50:45:10 v/v). After the coating with a isobutylmethacrate solution in hexane, the plate was incubated with the mAb NGR50 followed by a secondary HRP-conjugated anti-mouse IgG. The bands were identified using ECL. Band from each lane was scanned and quantitated using the ImageJ program.