Cholesterol detection kit

Manufactured by Abcam

Sourced in United States

The Cholesterol Detection Kit is a laboratory tool designed to quantitatively measure the level of cholesterol in a given sample. It provides a reliable and accurate method for determining the concentration of cholesterol present in various biological samples, such as blood, serum, or plasma.

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Lab products found in correlation

Application suite las, by Leica (1 mentions) Protein a g beads, by GE Healthcare (1 mentions) Ix71 microscope, by Hamamatsu Photonics (1 mentions) Orca ccd camera, by Hamamatsu Photonics (1 mentions) 96 well plate, by Corning (1 mentions)

4 protocols using cholesterol detection kit

Quantification of Intracellular Cholesterol and Neutral Lipids

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To determine the total intracellular cholesterol, the assay was carried out following the instructions of Cholesterol Detection Kit (cell-based) (BioVision Inc, San Francisco, USA). Briefly, cells were cultured with 50 and 70 μg/mL Yarrow for 48 hours before to be extracted with 200 μL of chloroform: Isopropanol: NP-40 (7:11:0.1) in a micro-homogenizer. The lipid fraction was collected and mingled with a cholesterol enzyme mix (which includes cholesterol esterase), and the absorbance of the resultant colorimetric assay was measured at 570 nm.
Quantification of neutral lipid content was done by mean of Bodipy staining. As a brief description, cells were treated with 2 μM BODIPY staining solution (BODIPY 493/503, Invitrogen) in PBS for 15 min at 37°C and fixed with 4% paraformaldehyde (PFA). Images were obtained with a Leica DM IL microscope, 40X Plan Fluotar objective and registered using Leica Application Suite (LAS).

Mouhid L., Gómez de Cedrón M., García-Carrascosa E., Reglero G., Fornari T, & Ramírez de Molina A. (2019). Yarrow supercritical extract exerts antitumoral properties by targeting lipid metabolism in pancreatic cancer. PLoS ONE, 14(3), e0214294.

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Quantification of MD-2-associated Cholesterol

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Five hundred µl of human LPDP was pre-cleared by incubation with 100 µl of protein A/G beads (GE Healthcare) for 2 hours at 4°C. After centrifugation, LPDP was incubated with 2 µg of either control rabbit IgG or rabbit anti-human MD-2 antibody [10 (link)] overnight at 4°C. Next day, protein A/G beads were added and incubated for 2 hours at 4°C. The samples were washed 5 times with PBS containing 0.1% Triton X-100 (PBS-T) and resuspended in 0.5 ml PBS. Cholesterol was extracted with 200 µl of chloroform:isopropanol:NP40 (7:11:0.1). After sonication and centrifugation, the organic phase was collected and dried out under argon. Unesterified cholesterol was measured using a cholesterol detection kit from BioVision. To verify cholesterol measurements, some samples, prior to cholesterol extraction and measurement, were cholesterol-depleted by incubation with 10 mM methyl-beta-cyclodextrin (MβCD) for 30 min at 37°C, washed 4 times with PBS-T and resuspended in 500 µl PBS. The same procedure was used to measure MD-2 associated cholesterol in mouse atherosclerotic lesions.

Choi S.H., Kim J., Gonen A., Viriyakosol S, & Miller Y.I. (2016). MD-2 binds cholesterol. Biochemical and biophysical research communications, 470(4), 877-880.

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Quantifying Cellular Cholesterol Levels

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The cells were seeded in 96-well plates (Corning Incorporated, NY, USA). Total cholesterol was visualized following the instruction on the cholesterol detection kit (cell-based) (Bio Vision, CA, USA). The images were obtained with an Olympus IX71 microscope using a Hamamatsu ORCA CCD camera.

Yu Q, & Cheng X. (2021). Hydroxyurea-induced membrane fluidity decreasing as a characterization of neuronal membrane aging in Alzheimer’s disease. Aging (Albany NY), 13(9), 12817-12832.

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Investigating LPCAT1 in Tumor Growth

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Six-week-old SCID/NOD mice were purchased from Jiangsu ALF Biotechnology Co., LTD (Nanjing, China). EC9706 and TE-1 cells (1 × 10⁷) transfected with lentivirus containing sh-LPCAT1 or sh-con were subcutaneously injected into the armpits of the mice. Tumor size and survival rate were measured every 7 days. On day 28, 5 mice each group were euthanized, and tumor tissues were harvested for cholesterol detection using a cholesterol detection kit (Biovision, USA). The survival time of each group of mice was recorded in another repeat experiment. This study was approved by the Ethics committee of Huai’an No. 1 People’s Hospital and was conducted in accordance with the guidelines of the National Animal Care and Ethics Institution [53 (link)].

Tao M., Luo J., Gu T., Yu X., Song Z., Jun Y., Gu H., Han K., Huang X., Yu W., Sun S., Zhang Z., Liu L., Chen X., Zhang L., Luo C, & Wang Q. (2021). LPCAT1 reprogramming cholesterol metabolism promotes the progression of esophageal squamous cell carcinoma. Cell Death & Disease, 12(9), 845.

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