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Cholesterol Uptake Assay Kit (ab236212) provides a convenient tool for studying cellular cholesterol trafficking.

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Functional Studies - Cholesterol Uptake Assay Kit (AB236212), expandable thumbnail
  • Functional Studies - Cholesterol Uptake Assay Kit (AB236212), expandable thumbnail
  • Functional Studies - Cholesterol Uptake Assay Kit (AB236212), expandable thumbnail

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Key facts

Detection method

Fluorescent

Sample types

Suspension cells, Adherent cells

Associated Products

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What's included?

1 Kit
Components
Cell-Based Assay Buffer Tablet
1 x 1 Tablet
Cell-Based Assay NBD Cholesterol
1 x 500 µL
Cell-Based Assay U-18666A
1 x 100 µL

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Cholesterol Uptake Assay Kit (ab236212) provides a convenient tool for studying cellular cholesterol trafficking.

Key facts

Detection method

Fluorescent

Sample types

Suspension cells, Adherent cells

Assay Platform

Microplate reader, Fluor. microscope, Flow cyt.

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage conditions

Multi

Appropriate long-term storage conditions

Multi

Storage information

Please refer to protocols

Notes

Cholesterol Uptake Assay Kit (ab236212) provides a convenient tool for studying cellular cholesterol trafficking. The kit employs NBD Cholesterol, a fluorescently-tagged cholesterol, as a probe for the detection of cholesterol taken up by cultured cells. U-18666A, which increases cholesterol uptake by inhibiting trafficking of synthesized cholesterol, is included as a positive control. The kit provides enough NBD Cholesterol to test 250 samples in a 96-well format.

Other cholesterol assay kits include:
- HDL and LDL/VLDL Cholesterol assay kit Cholesterol Assay Kit - HDL and LDL/VLDL ab65390
- Cell-based Cholesterol assay kit Cholesterol Assay Kit (Cell-Based) ab133116
- Cholesterol/Cholesterol Ester assay kit Cholesterol/ Cholesteryl Ester Assay Kit - Quantitation ab65359
- Cholesterol Efflux assay kit Cholesterol Efflux Assay Kit (Cell-based) ab196985

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

Cholesterol Uptake also referred to as cholesterol absorption involves key proteins like Niemann-Pick C1-Like 1 (NPC1L1). NPC1L1 is an important mediator of cholesterol transport across the intestinal epithelial cells. The protein has an approximate molecular weight of 145 kDa and is primarily expressed in the epithelial cells of the small intestine and in the liver. Its expression is regulated by various factors including dietary cholesterol levels which influences how it mediates cholesterol uptake into cells.

Biological function summary

Cholesterol uptake plays a significant role in maintaining cellular cholesterol homeostasis. The process involves NPC1L1 working as part of a larger macromolecular complex with other proteins like clathrin and adaptor protein 2 (AP2) which facilitate endocytosis of cholesterol into enterocytes. These interactions occur in the brush border of the intestinal lumen allowing for cholesterol to be transported and re-esterified into chylomicrons which then enter lymphatic circulation.

Pathways

Cholesterol uptake is a critical component of cholesterol metabolism and homeostasis. It is specifically involved in the reverse cholesterol transport pathway which manages the movement of cholesterol from peripheral tissues back to the liver. Within this pathway NPC1L1 interacts with other proteins such as apolipoprotein B48 (ApoB48) to facilitate the formation of chylomicrons that transport cholesterol. Another pathway influenced by cholesterol uptake is the bile acid synthesis pathway where excess cholesterol is converted to bile acids.

Associated diseases and disorders

Cholesterol uptake is connected to hypercholesterolemia and cardiovascular disease. Altered function or expression of NPC1L1 can result in variations in cholesterol levels potentially leading to the buildup of cholesterol in the bloodstream. This accumulation is a risk factor for cardiovascular diseases like atherosclerosis. Moreover inhibitors of NPC1L1 such as ezetimibe are used therapeutically to manage high cholesterol levels and prevent associated cardiovascular risks by decreasing intestinal cholesterol absorption.

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3 product images

  • Functional Studies - Cholesterol Uptake Assay Kit (ab236212), expandable thumbnail

    Functional Studies - Cholesterol Uptake Assay Kit (ab236212)

    U-18666A increases cholesterol uptake in Jurkat cells as measured by flow cytometry.

    Jurkat cells were seeded at a density of 5x105 cells/mL and incubated overnight in serum-free RPMI with U-18666A or vehicle and 20 μg/mL NBD Cholesterol in a cell culture incubator at 37°C. The next day, cells were transferred to a v-bottom plate for washing and reading on a flow cytometer. Cholesterol uptake was evaluated in the live cell gate using FlowJo analysis software. U-18666A at both 2.5 μM (green) and 1.25 μM (blue) showed a significant (p<0.05, t-test) shift in mean fluorescence as compared to the vehicle control (black) and the untreated (red) cells.

  • Functional Studies - Cholesterol Uptake Assay Kit (ab236212), expandable thumbnail

    Functional Studies - Cholesterol Uptake Assay Kit (ab236212)

  • Functional Studies - Cholesterol Uptake Assay Kit (ab236212), expandable thumbnail

    Functional Studies - Cholesterol Uptake Assay Kit (ab236212)

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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