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Cholesterol Assay Kit (Cell-Based) (ab133116)

  • Datasheet
  • SDS
  • Protocol Booklet
Reviews (1) Submit a question References (19)

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Cholesterol Assay Kit (Cell-Based) (ab133116)
  • Cholesterol Assay Kit (Cell-Based) (ab133116)
  • Cholesterol Assay Kit (Cell-Based) (ab133116)
  • Cholesterol Assay Kit (Cell-Based) (ab133116)
  • Cholesterol Assay Kit (Cell-Based) (ab133116)
  • Cholesterol Assay Kit (Cell-Based) (ab133116)

Key features and details

  • Assay type: Cell-based
  • Detection method: Fluorescent
  • Platform: Fluorescence microscope
  • Assay time: 1 hr 15 min
  • Sample type: Adherent cells

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Overview

  • Product name

    Cholesterol Assay Kit (Cell-Based)
    See all Cholesterol kits
  • Detection method

    Fluorescent
  • Sample type

    Adherent cells
  • Assay type

    Cell-based
  • Assay time

    1h 15m
  • Species reactivity

    Reacts with: Mammals, Other species
  • Product overview

    Cholesterol Assay Kit (Cell-Based) (ab133116) includes filipin III, fixative, and wash buffer in a ready to use format. It provides a simple fluorometric method to study mechanisms and biological factors that regulate cholesterol metabolism or movement within cells. A cholesterol trafficking inhibitor, U-18666A, is included as a positive control.


    Cholesterol assay protocol summary:
    - remove medium from cells
    - fix with fixative solution for 10 min
    - wash with wash buffer 3 times for 5 min
    - add Filipin III solution and incubate for 30-60 min
    - wash twice for 5 min
    - analyze with fluorescent microscope

  • Notes

    Cholesterol is both an important structural component of cell membranes and an early intermediate in hormone and bile acid biosynthesis. Cholesterol is not uniformly distributed among cellular membranes /  organelle membranes, but rather there are structurally and kinetically distinct cholesterol rich and poor domains.

    Filipin III is the predominant isomer of filipin, the collective name given to four isomeric polyene macrolides isolated from cultures of S. filipinensis. Filipin is widely used as a probe for sterol location in biological membranes.

    Interaction with cholesterol alters the filipin absorption and fluorescence spectra allowing visualization with a fluorescence microscope capable of excitation at 340-380 nm and emission at 385-470 nm. Filipin’s ease of use makes it a convenient tool for the histochemical identification of unesterified cholesterol both in vitro and in vivo.

    Other cholesterol assay kits include:
    - HDL and LDL/VLDL Cholesterol assay kit ab65390 
    - Cholesterol/Cholesterol Ester assay kit ab65359 
    - Cholesterol Efflux assay kit ab196985 
    - Cholesterol Uptake assay kit ab236212

    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

    Fluorescence microscope

Properties

  • Storage instructions

    Please refer to protocols.
  • Components 192 tests
    Cell-Based Assay Fixative 2 vials
    Cell-Based Assay U-18666A 1 vial
    Cholesterol Detection Assay Buffer 1 vial
    Cholesterol Detection Filipin III 1 vial
    Cholesterol Detection Wash Buffer 1 vial
  • Research areas

    • Cardiovascular
    • Lipids / Lipoproteins
    • Lipid Metabolism
    • Cholesterol Metabolism
    • Kits/ Lysates/ Other
    • Kits
    • Cell Metabolism Kits
    • Lipid Metabolism Kits
    • Cholesterol
    • Kits/ Lysates/ Other
    • Kits
    • Cell Metabolism Kits
    • Lipid Metabolism Kits
    • Obesity
    • Kits/ Lysates/ Other
    • Kits
    • Cell Metabolism Kits
    • Lipid Metabolism Kits
    • Lipid metabolism
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Lipid and lipoprotein metabolism
    • Cholesterol Metabolism
  • Relevance

    Cholesterol is a waxy substance that occurs naturally in all parts of the body and that the body needs to function normally. It is present in cell walls or membranes everywhere in the body, including the brain, nerves, muscle, skin, liver, intestines, and heart. The body uses cholesterol to produce many hormones, vitamin D, and the bile acids that help to digest fat. It takes only a small amount of cholesterol in the blood to meet these needs. However, if you have too much cholesterol in your bloodstream, it can lead to atherosclerosis, a condition in which fat and cholesterol are deposited in the walls of the arteries in many parts of the body, including the coronary arteries feeding the heart. In time, narrowing of the coronary arteries by atherosclerosis can produce the signs and symptoms of heart disease, including angina and heart attack.

Associated products

  • Related Products

    • Cholesterol/Cholesteryl Ester Assay Kit - Detection (ab102515)
    • Cholesterol/ Cholesteryl Ester Assay Kit - Quantitation (ab65359)

Images

  • Cholesterol Assay Kit (Cell-Based) (ab133116)
    Cholesterol Assay Kit (Cell-Based) (ab133116)

    HepG2 cells in response to 2 µM U-18666A.
    HepG2 cells were seeded in a 96-well plate at a density of 2.4 x 104 cells/well and cultured overnight. The next day, cells were treated with 2 µM U-18666A for 48 hours. U-18666A treatment for 48 hours induces intracellular accumulation.

  • Cholesterol Assay Kit (Cell-Based) (ab133116)
    Cholesterol Assay Kit (Cell-Based) (ab133116)

    HepG2 cells in response to 1 µM U-18666A.
    HepG2 cells were seeded in a 96-well plate at a density of 2.4 x 104 cells/well and cultured overnight. The next day, cells were treated with 1 µM U-18666A for 48 hours. U-18666A treatment for 48 hours induces intracellular accumulation.

     

  • Cholesterol Assay Kit (Cell-Based) (ab133116)
    Cholesterol Assay Kit (Cell-Based) (ab133116)

    HepG2 cells in response to 0.5 µM U-18666A.
    HepG2 cells were seeded in a 96-well plate at a density of 2.4 x 104 cells/well and cultured overnight. The next day, cells were treated with 0.5 µM U-18666A for 48 hours. U-18666A treatment for 48 hours induces intracellular accumulation.

  • Cholesterol Assay Kit (Cell-Based) (ab133116)
    Cholesterol Assay Kit (Cell-Based) (ab133116)

    HepG2 cells in response to 0.25 µM U-18666A.
    HepG2 cells were seeded in a 96-well plate at a density of 2.4 x 104 cells/well and cultured overnight. The next day, cells were treated with 0.25 µM U-18666A for 48 hours. U-18666A treatment for 48 hours induces intracellular accumulation.

     

  • Cholesterol Assay Kit (Cell-Based) (ab133116)
    Cholesterol Assay Kit (Cell-Based) (ab133116)

    Accumulation of cholesterol inside HepG2 cells - Control.
    HepG2 cells were seeded in a 96-well plate at a density of 2.4 x 104 cells/well and cultured overnight. The next day, cells were treated with DMSO (vehicle) for 48 hours. Cells demonstrate that majority of cholesterol is localized on the plasma membrane.

  • Cholesterol Assay Kit (Cell-Based) (ab133116)
    Cholesterol Assay Kit (Cell-Based) (ab133116)
    Accumulation of cholesterol inside HepG2 cells in response to 1.25 µM U-18666A.
    HepG2 cells were seeded in a 96-well plate at a density of 3 x 104 cells/well and cultured overnight. The next day, cells were treated with DMSO (vehicle) or 1.25 µM U-18666A for 48 hours. Panel A: Cells treated with DMSO alone demonstrate that majority of cholesterol is localized on the plasma membrane. Panel B: U-18666A treatment for 48 hours induces intracellular accumulation.

Protocols

  • Protocol Booklet

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (19)

Publishing research using ab133116? Please let us know so that we can cite the reference in this datasheet.

ab133116 has been referenced in 19 publications.

  • Triplet EM  et al. The thrombin receptor links brain derived neurotrophic factor to neuron cholesterol production, resiliency and repair after spinal cord injury. Neurobiol Dis 152:105294 (2021). PubMed: 33549720
  • Nadeem A  et al. Phosphatidic acid-mediated binding and mammalian cell internalization of the Vibrio cholerae cytotoxin MakA. PLoS Pathog 17:e1009414 (2021). PubMed: 33735319
  • Jin Y  et al. A Novel c-MET-Targeting Antibody-Drug Conjugate for Pancreatic Cancer. Front Oncol 11:634881 (2021). PubMed: 33816276
  • Shin S  et al. Qki activates Srebp2-mediated cholesterol biosynthesis for maintenance of eye lens transparency. Nat Commun 12:3005 (2021). PubMed: 34021134
  • Sun X  et al. miR-344-5p Modulates Cholesterol-Induced ß-Cell Apoptosis and Dysfunction Through Regulating Caveolin-1 Expression. Front Endocrinol (Lausanne) 12:695164 (2021). PubMed: 34394002
View all Publications for this product

Customer reviews and Q&As

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Accumulation of cholesterol in WT mouse embryonic limb mesenchymal cells

Excellent Excellent 5/5 (Ease of Use)
Abreviews
Abreviews
abreview image
Cells were seeded in 96-well plate and cultured for 36h. Experiment has been performed by Julie Gamart, PhD.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Dr. Julie Gamart

Verified customer

Submitted Apr 17 2018

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