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.
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- Scientific reports 13:197252023Induced alternative splicing an opportunity to study PCSK9 protein isoforms at physiologically relevant concentrations.Applications:Unspecified applicationReactive species:Unspecified reactive speciesJessica M Cale,Kristin A Ham,Dunhui Li,Craig S McIntosh,Gerald F Watts,Steve D Wilton,May T Aung-HtutPubMed 37957262
- ACS omega 8:30906-309162023Triterpenoids from Exhibit Hypocholesterolemic Potential via Regulation of Cholesterol Biosynthesis and Stimulation of Low-Density Lipoprotein Uptake in HepG2 Cells.Applications:Unspecified applicationReactive species:Unspecified reactive speciesMusawenkosi Ndlovu,June C Serem,Mamoalosi A Selepe,Andrew R Opoku,Megan J Bester,Zeno Apostolides,Rebamang A MosaPubMed 37663489
- iScience 24:1031802021NRF3 upregulates gene expression in SREBP2-dependent mevalonate pathway with cholesterol uptake and lipogenesis inhibition.Applications:Unspecified applicationReactive species:Unspecified reactive speciesTsuyoshi Waku,Toru Hagiwara,Natsuko Tamura,Yuri Atsumi,Yasuomi Urano,Mikiko Suzuki,Takuya Iwami,Katsuya Sato,Masayuki Yamamoto,Noriko Noguchi,Akira KobayashiPubMed 34667945
Key facts
- Detection method
- Flow cytometry-fluorescent
- Sample types
- Adherent cells
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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.
Key facts
- Detection method
- Flow cytometry-fluorescent
- Sample types
- Adherent cells
- Assay Platform
- Flow cytometer
Storage
- Shipped at conditions
- Blue Ice
- Appropriate short-term storage conditions
- +4°C
- Appropriate long-term storage conditions
- Multi
- Storage information
- Please refer to protocols
Notes
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.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
Low-density lipoprotein often referred to as LDL or "bad cholesterol" serves as a carrier for lipids in the bloodstream. LDL particles are spherical and contain an apolipoprotein B-100 shell with a core mostly filled with cholesterol esters. It has an approximate molecular weight of 2536 kDa. LDL is mainly expressed in the liver where it is released into the circulation to deliver cholesterol to various tissues. It is an essential component that transports cholesterol and other lipids through the body for structural and metabolic functions.
- Biological function summary
LDL plays an important role by supplying cholesterol to cells which is critical for membrane structure and hormone synthesis. LDL binds to specific receptors on the cell surface prompting its cellular uptake through the LDL uptake assay mechanisms including receptor-mediated endocytosis. This process involves the LDL protein engaging with a receptor complex facilitating the internalization of cholesterol for cell usage or storage. Native LDL can be distinguished from oxidized forms which have different roles in health and disease.
- Pathways
LDL is vital in cholesterol homeostasis and lipid metabolism pathways. One notable pathway includes its interaction in the mevalonate pathway essential for cholesterol biosynthesis. The interaction with the insulin signaling pathway also highlights the significance of LDL in energy homeostasis. Related proteins such as high-density lipoprotein (HDL) and very low-density lipoprotein (VLDL) work in tandem with LDL forming a complex system for managing lipid transport and affecting cholesterol levels.
- Associated diseases and disorders
LDL is closely associated with atherosclerosis and cardiovascular disease. High LDL levels lead to cholesterol accumulation in arterial walls promoting the formation of plaques and increasing the risk of heart disease. Apolipoprotein B-100 part of the LDL structure is often investigated in the context of these diseases because it mediates LDL's attachment to arterial walls. Understanding LDL's relationship with these disorders highlights the importance of controlling its levels through medical and lifestyle interventions therefore minimizing disease risk.
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Functional Studies - LDL Uptake Assay Kit (Flow cytometry) (ab236208)
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.
Functional Studies - LDL Uptake Assay Kit (Flow cytometry) (ab236208)
Lovastatin increases uptake of LDL.
HepG2 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.
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