LDL Uptake Assay Kit (Cell-Based) (ab133127)
Key features and details
- Assay type: Cell-based
- Detection method: Fluorescent
- Platform: Fluorescence microscope
- Assay time: 6 hr 30 min
- Sample type: Adherent cells
Overview
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Product name
LDL Uptake Assay Kit (Cell-Based) -
Detection method
Fluorescent -
Sample type
Adherent cells -
Assay type
Cell-based -
Assay time
6h 30m -
Species reactivity
Reacts with: Mammals, Other species -
Product overview
LDL Uptake Assay Kit (Cell-Based) (ab133127) provides a convenient tool for studying LDL uptake and regulation at the cellular level. This kit employs Human LDL conjugated to DyLight™ 550 as a fluorescent probe for detection of LDL uptake into cultured cells. An LDL receptor-specific polyclonal antibody and a DyLight™ 550-conjugated secondary antibody are included for identifying the distribution of LDL receptors.
LDL uptake assay protocol summary:
- remove culture medium from experimentally treated cells
- add LDL-Dylight 550 solution and incubate for 3-4 hrs
- replace solution with culture medium
- analyze LDL uptake with fluorescent microscope
- wash cells and fix for 10 min
- wash 3 times for 5 min
- add assay blocking solution and incubate for 30 min
- add anti-LDL receptor antibody and incubate for 1 hr
- wash 3 times for 5 min
- add Dylight 488 secondary antibody and incubate for 1 hr
- wash 3 times for 5 min
- analyze staining with fluorescence microscope -
Notes
LDL is the major carrier of cholesterol in the blood, accounting for more than 60% of total plasma cholesterol. LDL is taken up by hepatic and extra-hepatic tissues through receptor mediated endocytosis triggered by apoB-100-LDL receptor interaction. The internalized LDL particle is transported to lysosomes where it is degraded to free cholesterol and amino acids.
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Platform
Fluorescence microscope
Properties
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Storage instructions
Please refer to protocols. -
Components 1 kit Cell-Based Assay Blocking Solution 1 vial Cell-Based Assay Fixative 1 vial DyLight 488-Conjugated Goat Anti-Rabbit Secondary Antibody 1 unit LDL-DyLight 550 1 vial Rabbit Anti-LDL Receptor Primary Antibody 1 vial -
Research areas
Images
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Functional Studies - LDL Uptake Assay Kit (Cell-Based) (ab133127)LDL uptake in pre-adipocytes and adipocytes.
Panel A: Light image of undifferentiated pre-adipocytes.
Panel B: Undifferentiated pre-adipocytes do not show any LDL uptake.
Panel C: Undifferentiated pre-adipocytes show little LDL receptor expression.
Panel D: Light image of differentiated adipocytes.
Panel E: Corresponding cells to those in Panel D showing uptake of LDL (red).
Panel F: Corresponding cells to those in Panel D showing expression of LDL receptor (green). -
LDL Uptake Assay Kit (Cell-Based) (ab133127)LDL uptake in HepG2 cells.
HepG2 cells were cultured at a density of 3 x 104 cells/well in a 96 well plate for two days then treated with 32.5µM EGCG overnight. LDL-Dylight™ 549 (10µg/ml) was added and the cells were incubated for an additional four hours. Cells were fixed and stained for LDL receptor using a Rabbit anti-LDL receptor primary antibody and DylightTM 488-conjugated secondary antibody.
Panel A: LDL-Dylight™ 549 taken into cells appears in red.
Panel B: LDL receptors (in green) show a distribution pattern that matches the cells in the Panel A containing LDL-Dylight™ 549.
