Phospholipid Synthesis Assay Kit (ab241025)
Overview
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Product name
Phospholipid Synthesis Assay Kit
See all Phospholipid kits -
Product overview
The Phospholipid Synthesis Assay Kit (ab241025) offers a simple and robust method to label and visualize newly synthesized phospholipids in vivo. Based on the metabolic incorporation of the choline analogs directly into their structure, modified phospholipid molecules can be detected with high sensitivity and spatial resolution by click chemistry with azide-containing dyes.
This kit enables analyses of global biosynthesis, subcellular localization and turnover of Cho-containing phospholipids in cells. Cells show strong incorporation of Cho analogs into all classes of phospholipids that can be assayed by fluorescence microscopy, or quantified by FACS.
Properties
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Storage instructions
Store at -20°C. Please refer to protocols. -
Components 100 tests Copper Reagent (100X) 1 x 100µl Fixative Solution 1 x 10ml Fluorescent Azide (100X) 1 x 100µl Permeabilization Buffer (10X) 1 x 25ml Phospholipid Label (1000X) 1 x 10µl Reducing Agent (20X) 1 x 500µl Total DNA Stain (1000X) 1 x 10µl Wash Buffer (10X) 1 x 25ml
Images
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Analysis of metabolic labeling of phospholipids in proliferating cells.
Jurkat cells (1X106 cells/ml) were pre-treated with vehicle (black line) or cultured in presence of 1X Phospholipid Label (red line) for 24 hours at 37°C prior to 1 hour incubation with Phospholipase D (blue line). Modified phospholipid molecules were detected according to the kit protocol and red fluorescence was analyzed by FACS in FL-2 channel. Decrease in signal caused by hydrolysis of Cho-containing head groups via Phospholipase D activity confirms that red fluorescence is the result of Phospholipid Label incorporation.
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Analysis of metabolic labeling of phospholipids in proliferating cells.BALB/3T3 cells (105 cells/ ml) cultured in presence of ab241025 for 24 hours at 37°C and processed according to kit protocol. Choline-containing phospholipids were detected by Fluorescence Microscope according to kit protocol.
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Analysis of metabolic labeling of phospholipids in proliferating cells.Total DNA staining confirms that red fluorescence is the result of Phospholipid Label incorporation.
