Cholesterol Assay Kit (Cell-Based) (ab133116) includes filipin III, fixative, and wash buffer in a ready to use format.
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- npj aging 10:52024Cholesterol biosynthetic pathway induces cellular senescence through ERRα.Applications:Unspecified applicationReactive species:Unspecified reactive speciesDorian V Ziegler,Joanna Czarnecka-Herok,Mathieu Vernier,Charlotte Scholtes,Clara Camprubi,Anda Huna,Amélie Massemin,Audrey Griveau,Christelle Machon,Jérôme Guitton,Jennifer Rieusset,Arnaud M Vigneron,Vincent Giguère,Nadine Martin,David BernardPubMed 38216569
- iScience 26:1081442023Deciphering the mechanism of action of VP343, an antileishmanial drug candidate, in .Applications:Unspecified applicationReactive species:Unspecified reactive speciesSameh Obeid,Eloisa Berbel-Manaia,Valérie Nicolas,Indira Dennemont,Julien Barbier,Jean-Christophe Cintrat,Daniel Gillet,Philippe M Loiseau,Sébastien PomelPubMed 37915600
- Nature communications 14:51502023Astroglial exosome HepaCAM signaling and ApoE antagonization coordinates early postnatal cortical pyramidal neuronal axon growth and dendritic spine formation.Applications:Unspecified applicationReactive species:Unspecified reactive speciesShijie Jin,Xuan Chen,Yang Tian,Rachel Jarvis,Vanessa Promes,Yongjie YangPubMed 37620511
- Stem cell research & therapy 14:2142023APOE deficiency impacts neural differentiation and cholesterol biosynthesis in human iPSC-derived cerebral organoids.Applications:Unspecified applicationReactive species:Unspecified reactive speciesJing Zhao,Tadafumi C Ikezu,Wenyan Lu,Jesse R Macyczko,Yonghe Li,Laura J Lewis-Tuffin,Yuka A Martens,Yingxue Ren,Yiyang Zhu,Yan W Asmann,Nilüfer Ertekin-Taner,Takahisa Kanekiyo,Guojun BuPubMed 37605285
- British journal of cancer 128:1991-19992023MBTPS2 acts as a regulator of lipogenesis and cholesterol synthesis through SREBP signalling in prostate cancer.Applications:Unspecified applicationReactive species:Unspecified reactive speciesAmy J Tibbo,Andrew Hartley,Richa Vasan,Robin Shaw,Laura Galbraith,Ernest Mui,Hing Y Leung,Imran AhmadPubMed 36991255
- Cancer cell 41:740-756.e102023Targeting the mevalonate pathway suppresses ARID1A-inactivated cancers by promoting pyroptosis.Applications:Unspecified applicationReactive species:Unspecified reactive speciesWei Zhou,Heng Liu,Zhe Yuan,Joseph Zundell,Martina Towers,Jianhuang Lin,Simona Lombardi,Hao Nie,Brennah Murphy,Tyler Yang,Chen Wang,Liping Liao,Aaron R Goldman,Toshitha Kannan,Andrew V Kossenkov,Ronny Drapkin,Luis J Montaner,Daniel T Claiborne,Nan Zhang,Shuai Wu,Rugang ZhangPubMed 36963401
- FASEB journal : official publication of the Federation of American Societies for Experimental Biology 37:e227152022Annexin A2 modulates phospholipid membrane composition upstream of Arp2 to control angiogenic sprout initiation.Applications:Unspecified applicationReactive species:Unspecified reactive speciesTimothy M Sveeggen,Colette A Abbey,Rebecca L Smith,Michael L Salinas,Robert S Chapkin,Kayla J BaylessPubMed 36527391
- iScience 25:1054072022Citrate shuttling in astrocytes is required for processing cocaine-induced neuron-derived excess peroxidated fatty acids.Applications:Unspecified applicationReactive species:Unspecified reactive speciesKalimuthusamy Natarajaseenivasan,Alvaro Garcia,Prema Velusamy,Santhanam Shanmughapriya,Dianne LangfordPubMed 36389000
- Cancer research 82:3845-38572022Inhibition of EGFR Overcomes Acquired Lenvatinib Resistance Driven by STAT3-ABCB1 Signaling in Hepatocellular Carcinoma.Applications:Unspecified applicationReactive species:Unspecified reactive speciesBeiyuan Hu,Tiantian Zou,Wei Qin,Xiaotian Shen,Yinghan Su,Jianhua Li,Yang Chen,Ze Zhang,Haoting Sun,Yan Zheng,Chao-Qun Wang,Zhengxin Wang,Tian-En Li,Shun Wang,Le Zhu,Xufeng Wang,Yan Fu,Xudong Ren,Qiongzhu Dong,Lun-Xiu QinPubMed 36066408
- Journal of advanced research 45:1-132022SCAP deficiency facilitates obesity and insulin resistance through shifting adipose tissue macrophage polarization.Applications:Unspecified applicationReactive species:Unspecified reactive speciesJae-Ho Lee,Sun Hee Lee,Eun-Ho Lee,Jeong-Yong Cho,Dae-Kyu Song,Young Jae Lee,Taeg Kyu Kwon,Byung-Chul Oh,Kae Won Cho,Timothy F Osborne,Tae-Il Jeon,Seung-Soon ImPubMed 35659922
Key facts
- Detection method
- Fluorescent
- Sample types
- Adherent cells
- Assay type
- Cell-based
- Reactive species
- Mammals
- Assay time
- 1h 15m
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Cholesterol Assay Kit (Cell-Based) (ab133116) includes filipin III, fixative, and wash buffer in a ready to use format.
Key facts
- Detection method
- Fluorescent
- Sample types
- Adherent cells
- Assay type
- Cell-based
- Reactive species
- Mammals
- Assay time
- 1h 15m
- Assay Platform
- Fluorescence microscope
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 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 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.
How the cell-based cholesterol assay works
Cholesterol Assay Kit (Cell-Based) includes filipin III, fixative, and wash buffer in a ready to use format.
Filipin III is the predominant isomer of filipin, the collective name given to four isomeric polyene macrolides isolated from cultures of S.filipinensis.
Filipin has been 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.
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
Other cholesterol assay kits include:
- HDL and LDL/VLDL Cholesterol assay kit Cholesterol Assay Kit - HDL and LDL/VLDL ab65390
- Cholesterol/Cholesterol Ester assay kit Cholesterol/ Cholesteryl Ester Assay Kit - Quantitation ab65359
- Cholesterol Efflux assay kit Cholesterol Efflux Assay Kit (Cell-based) ab196985
- Cholesterol Uptake assay kit 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.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
Cholesterol sometimes referred to as fchol is a lipid molecule with a molecular mass of approximately 386.65 Daltons. It is an essential component of cell membranes and is found in the plasma membranes of cells throughout the body. Cholesterol primarily resides in the membranes where it modulates fluidity and mechanical properties. Its expression occurs in various tissues but it is abundant in the liver which plays an important role in its synthesis and regulation.
- Biological function summary
Cholesterol is important in maintaining the structural integrity and organization of membranes. It is also part of lipid rafts which are specialized microdomains within the cell membrane that facilitate cellular signaling. Cholesterol interacts with proteins and receptors embedded in the membrane influencing their function. It does not form a complex itself but affects complexes within the membrane like lipid rafts and signaling platforms.
- Pathways
Cholesterol is a critical component of the steroid biosynthesis pathway and the bile acid synthesis pathway. It serves as a precursor for steroid hormones including cortisol and sex hormones as well as bile acids which are essential for digestion and lipid absorption. Proteins such as HMG-CoA reductase and the low-density lipoprotein receptor (LDLR) are key players related to cholesterol in these pathways regulating its synthesis and uptake in cells.
- Associated diseases and disorders
Cholesterol is closely associated with atherosclerosis and cardiovascular disease. Elevated levels of low-density lipoprotein (LDL) cholesterol contribute to plaque formation in arteries increasing the risk of heart attack and stroke. On the other hand high-density lipoprotein (HDL) cholesterol helps remove excess cholesterol from tissues including the arterial walls. Statins a class of drugs that lower LDL cholesterol levels target HMG-CoA reductase and are commonly used in treating hypercholesterolemia and cardiovascular diseases.
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6 product images
Fluorescence Microscopy - 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.
Fluorescence Microscopy - 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.
Fluorescence Microscopy - 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.
Fluorescence Microscopy - 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.
Fluorescence Microscopy - 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.
Fluorescence Microscopy - 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.
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