Total Phospholipid Assay Kit (Cell-Based) (ab273294)
Key features and details
- Assay type: Cell-based
- Detection method: Fluorescent
- Platform: Microplate reader, Fluorescence microscope
- Sample type: Adherent cells, Suspension cells
Overview
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Product name
Total Phospholipid Assay Kit (Cell-Based)
See all Phospholipid kits -
Detection method
Fluorescent -
Sample type
Adherent cells, Suspension cells -
Assay type
Cell-based -
Assay duration
Multiple steps standard assay -
Product overview
Total Phospholipid Assay Kit (Cell-Based) (ab273294) 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 (Ex/Em= 494/521 nm). This kit enables quantitative analyses of global biosynthesis/turnover of Cho-containing phospholipids in cells. Cells show strong incorporation of Cho analogs into all classes of phospholipids that can be assayed by microplate reader and fluorescence microscope. The kit provides sufficient materials for 100 assays.
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Notes
Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses. -
Platform
Microplate reader, Fluorescence microscope
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 20µl Wash Buffer (10X) 1 x 25ml
Images
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Analysis of metabolic labeling of phospholipids in proliferating cells.
Jurkat cells (1X106 cells/well) were pre-treated with vehicle or cultured in presence of 1X Phospholipid Label for 24 hours at 37°C prior to 1 hour treatment with Phospholipase D and then processed for detection of according to the kit protocol. Plate reader analyses of controls and PLD treatment; Avg fluorescence +/- standard deviation plotted for 3 replicates per condition.
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Analysis of metabolic labeling of phospholipids in proliferating cells.BALB/3T3 cells seeded at 105 cells/ ml were pre-treated with vehicle or cultured in presence of 1X Phospholipid Label for 24 hours at 37°C prior to 1 hour treatment with Phospholipase D and then processed for detection of according to the kit protocol. BALB/3T3 cells: left panel- green fluorescence of de novo synthesized phospholipids; right panel-nuclear staining and phospholipid images merged.
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Analysis of metabolic labeling of phospholipids in proliferating cells.Jurkat cells (1X106 cells/well) were pre-treated with vehicle or cultured in presence of 1X Phospholipid Label for 24 hours at 37°C prior to 1 hour treatment with Phospholipase D and then processed for detection of according to the kit protocol. Fluorescence Azide Curve of Jurkat cells prepared for this assay. Detection limit corresponds to about 31,250 of Jurkat cells per well. Your results may not be identical to these. A new curve must be obtained for each experiment and the cell line.
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Azide Fluorescence Curve in 0-0.1 X range.
