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Acetylated ldl

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Acetylated LDL is a laboratory reagent used in research applications. It is a modified form of low-density lipoprotein (LDL) that has been chemically acetylated. This product is designed for use in cell-based assays and other research procedures where acetylated LDL is required.

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Lab products found in correlation

Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions) Nunc lab tek 2 cc2 chamber slide, by Thermo Fisher Scientific (2 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions) P4359, by Merck Group (2 mentions) Ulex lectin, by Merck Group (1 mentions) Amiloride, by Merck Group (1 mentions) 1 1 dioctadecyl 3 3 3 3 tetramethylindodicarbocyanine, by Thermo Fisher Scientific (1 mentions) 2 hydroxypropyl β cyclodextrin h107, by Merck Group (1 mentions) Endo h, by New England Biolabs (1 mentions) Pngase f, by New England Biolabs (1 mentions) Macrophage colony stimulating factor m csf, by Thermo Fisher Scientific (1 mentions) Penicillin streptomycin l glutamine, by Thermo Fisher Scientific (1 mentions)

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Acetylated low-density lipoprotein (ac-LDL; (2 citations)

6 protocols using acetylated ldl

1

Acetylated LDL Uptake Assay

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6×104 fibroblasts per cm2 were cultured overnight at 37°C, 5% CO2 on Growth Factor Reduced Matrigel Basement Membrane Matrix (BD) coated wells of Nunc Lab-Tek II CC2 chamber slides (Thermo). Serum starved or unstarved cells were cultured in IMDM, 1X Penicillin/Streptomycin or IMDM, 1X Penicillin/Streptomycin, 10% fetal bovine serum (Gibco), respectively. Pifithrin-α 100μM (P4359, Sigma) or RITA 0.1μM (506149, EMD Chemicals) were added to the cells cultured with the above culture medium. Acetylated LDL (Invitrogen) uptake was performed as described40 (link).
Ubil E., Duan J., Pillai I.C., Rosa-Garrido M., Wu Y., Bargiacchi F., Lu Y., Stanbouly S., Huang J., Rojas M., Vondriska T.M., Stefani E, & Deb A. (2014). Mesenchymal-endothelial-transition contributes to cardiac neovascularization. Nature, 514(7524), 585-590.
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2

Acetylated LDL Uptake Assay

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6×104 fibroblasts per cm2 were cultured overnight at 37°C, 5% CO2 on Growth Factor Reduced Matrigel Basement Membrane Matrix (BD) coated wells of Nunc Lab-Tek II CC2 chamber slides (Thermo). Serum starved or unstarved cells were cultured in IMDM, 1X Penicillin/Streptomycin or IMDM, 1X Penicillin/Streptomycin, 10% fetal bovine serum (Gibco), respectively. Pifithrin-α 100μM (P4359, Sigma) or RITA 0.1μM (506149, EMD Chemicals) were added to the cells cultured with the above culture medium. Acetylated LDL (Invitrogen) uptake was performed as described40 (link).
Ubil E., Duan J., Pillai I.C., Rosa-Garrido M., Wu Y., Bargiacchi F., Lu Y., Stanbouly S., Huang J., Rojas M., Vondriska T.M., Stefani E, & Deb A. (2014). Mesenchymal-endothelial-transition contributes to cardiac neovascularization. Nature, 514(7524), 585-590.
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3

Endothelial Progenitor Cell Enumeration

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CACs were the adherent mononuclear cells of a 4- to 7-day culture procedure (23 (link)). The number of CAC cells was assessed in 15 SCT and 14 control subjects. Mononuclear cells (1 × 106) were plated on fibronectin-coated 24-well plates enriched with endothelial medium (basal medium with supplement pack, PromoCell, Heidelberg, Germany, supplemented with 20% fetal calf serum). The medium was changed after 72 and 120 h.
Counting was performed on day 5 in 15 randomly selected high-power fields (HPF, ×400) by fluorescence microscopy after incubation with acetylated-LDL (5 mg/L; Invitrogen, Carlsbad, CA, USA) and Ulex lectin (10 mg/L; Sigma, Saint Louis, MO, USA) for 4 h at 37°C. CAC cells were identified as cells with combined uptake of acetylated-LDL and Ulex lectin binding.
Phowira J., Bakhashab S., Doddaballapur A, & Weaver J.U. (2022). Subclinical Thyrotoxicosis and Cardiovascular Risk: Assessment of Circulating Endothelial Progenitor Cells, Proangiogenic Cells, and Endothelial Function. Frontiers in Endocrinology, 13, 894093.
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4

Radiolabeled Cholesterol Efflux

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J774A.1 cells were incubated with 1μCi/mL 3H-labeled cholesterol and 50μg/mL acetylated LDL (Alfa Aesar, Tewksbury, MA) for 24 hours. Cells were washed and then exposed to serum-free RPMI ± PCM (± PGE2 antibody) overnight. Cells were washed with PBS and then exposed to 50μg/mL apoA1 (Alfa Aesar, Tewksbury, MA) for 4 hours. Liquid scintillation counting was then used to quantify the percentage of radiolabeled cholesterol present in the apoA-1-containing fraction.
Heffron S.P., Weinstock A., Scolaro B., Chen S., Sansbury B.E., Marecki G., Rolling C.C., El Bannoudi H., Barrett T., Canary J.W., Spite M., Berger J.S, & Fisher E.A. (2020). Platelet Conditioned Media Induces an Anti-inflammatory Macrophage Phenotype through EP4. Journal of thrombosis and haemostasis : JTH, 19(2), 562-573.
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5

Lipid Reagents for Membrane Biophysical Studies

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2-Hydroxypropyl-β-cyclodextrin (H-107) and amiloride (A7410) were from Sigma. EndoH (P0702) and PNGaseF (P0704) were from New England Biolabs. Dynasore (14061) was from Cayman Chemical; Human HDL (J64903) and acetylated LDL (J65029) were from Alfa Aesar. 5A peptide (DWLKAFYDKVAEKLKEAF-P-DWAKAAYDKAAEKAKEAA, 4078379) was from Bachem Americas (Torrance, CA). 22A peptide (PVLDLFRELLNELLEALKQKLK) was synthesized by Genscript (Piscataway, NJ). Lipids including egg-sphingomyelin (SM, Coastome NM-10), 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC, Coastome MC-4040), and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC, Coastome MC-6081) were from NOF America Corporation. 1,1′-Dioctadecyl-3,3,3′,3′-tetramethylindodicarbocyanine, 4-chlorobenzenesulfonate salt (DiD, D7757) and 4-(4-(dihexadecylamino)styryl)-N-methylpyridinium iodide (DiA, D3883) were from Invitrogen. Cholesteryl [1,2,6,7-3H(N)] linoleate (ART 1203) and sphingomyelin [choline methyl-3H] were from American Radiolabeled Chemicals (Saint Louis, MO). N-[6-[(7-Nitro-2-1,3-benxoxadiazol-4-yl)amino]-sphingosine-1-phosphocholine (NBD-Sphingomyelin, 810218P) was from Avanti Polar Lipids (Alabaster, AL).
Schultz M.L., Fawaz M.V., Azaria R.D., Hollon T.C., Liu E.A., Kunkel T.J., Halseth T.A., Krus K.L., Ming R., Morin E.E., McLoughlin H.S., Bushart D.D., Paulson H.L., Shakkottai V.G., Orringer D.A., Schwendeman A.S, & Lieberman A.P. (2019). Synthetic high-density lipoprotein nanoparticles for the treatment of Niemann–Pick diseases. BMC Medicine, 17, 200.
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6

Isolation and Polarization of Bone Marrow-Derived Macrophages

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Bone marrow-derived macrophages were isolated and cultured as follows. Briefly, 3-month-old mice were euthanized by carbon dioxide inhalation. Hind legs were exposed and the muscle tissue removed to isolate the femurs and tibiae. The bone marrow was flushed from femurs and tibiae with culture medium (DMEM) containing 10% fetal calf serum and 1% penicillin/streptomycin/l-glutamine (Invitrogen, Carlsbad, CA) using a syringe and a 25-gauge needle. The cell suspension was drawn through a syringe with an 18-gauge needle to dissociate cell clumps and was passed through a 70-μm pore cell strainer (BD BioSciences, San Jose, CA) to remove tissue debris. The cells were plated and incubated for 5 days in the presence of 25 ng/ml macrophage colony stimulating factor (M-CSF) (PeproTech, Rocky Hill, NJ) in order to initiate macrophage differentiation. Foam cell differentiation from macrophages was achieved with 50 μg/ml acetylated LDL (Alfa Aesar, J65029) added to the cell culture media for 48 h. For macrophage polarization towards the classic M1 and alternate M2 phenotypes, macrophages were kept for 24 h in DMEM medium supplemented with 20 ng/mL TNFα for M1 differentiation and with 20 ng/mL IL-4 for M2 differentiation. Cells were harvested after 24 h for total RNA isolation.
Brampton C., Pomozi V., Chen L.H., Apana A., McCurdy S., Zoll J., Boisvert W.A., Lambert G., Henrion D., Blanchard S., Kuo S., Leftheriotis G., Martin L, & Le Saux O. (2021). ABCC6 deficiency promotes dyslipidemia and atherosclerosis. Scientific Reports, 11, 3881.
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