LDL Uptake Assay Kit (Flow cytometry) (ab236208)
Key features and details
- Detection method: Flow cytometry-fluorescent
- Platform: Flow cytometer
- Sample type: Adherent cells
Overview
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Product name
LDL Uptake Assay Kit (Flow cytometry) -
Detection method
Flow cytometry-fluorescent -
Sample type
Adherent cells -
Product overview
The LDL Uptake Assay Kit (Flow cytometry) (ab236208) employs human LDL conjugated to DyLightTM 488 as a convenient tool for studying the uptake of LDL in cultured cells. Flow cytometry provides the advantage of assessing the uptake of LDL at the single-cell level. In addition, multiplexing with other markers, such as LDLR expression, is possible to gain more information from a single experiment. The reagents provided in this kit are sufficient to test 48 samples by flow cytometry.
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Platform
Flow cytometer
Properties
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Storage instructions
Store at +4°C. Please refer to protocols. -
Components 1 x 48 tests Cell-Based Assay 7-AAD Staining Stock Solution (1000X) 1 x 50µl Cell-Based Assay Buffer Tablet 1 tablet LDL-DyLightTM 488 Assay Reagent 1 x 120µl Lovastatin Control 1 x 0.1mg -
Research areas
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Relevance
The low density lipoprotein (LDL) receptor system coordinates the metabolism of cholesterol, an essential component of the plasma membrane of all mammalian cells. Study of this system has led to an enhanced understanding of the cellular basis of cholesterol homeostasis. It has also brought into focus an important mechanism of metabolic regulation--the process of receptor-mediated endocytosis (1). Data suggest that the juxtamembranous region of the cytoplasmic domain participates in protein:protein interactions that allow the low density lipoprotein receptor to cluster in coated pits (2). It has been shown that the family of LDL receptors may serve as viral receptors. Endocytosis of the Flaviviridae viruses, hepatitis C virus, GB virus C/hepatitis G virus, and bovine viral diarrheal virus (BVDV) was shown to be mediated by LDL receptors on cultured cells -
Cellular localization
Cell Membrane -
Alternative names
- low density lipoprotein
Images
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Example flow cytometric analysis
HepG2 cells were plated at 2 x 105 cells/well in a 24 well plate approximately 48 hours before addition of LDL-DyLightTM 488. After a four hour incubation with the LDL probe, cells were trypsinized, washed and stained with 7-AAD prior to flow cytometry. In this example analysis, after digital compensation, 7-AAD negative live cells are gated first, followed by scatter. Single cells are gated using an area versus height dot plot, and the geometric mean fluorescence intensity (MFI) of the resulting cells in the LDL-DyLightTM 488 channel is determined.
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Lovastatin increases uptake of LDLHepG2 cells were plated at 2 x 105 cells/well in a 24 well plate and allowed to adhere overnight, before being treated with 1 µM Lovastatin or left untreated for 24 hours in MEM + 2% FBS. The final four hours of treatment included the probe LDL-DyLightTM 488. Cells were processed as described in the kit booklet and the flow cytometric data were analyzed as described in Figure 1. Average geometric mean fluorescence intestities (MFI) for each group are plotted.
