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Lipoprotein preparation

Lipoprotein preparation - Protocols and Methods Forum

Lipoprotein preparation - Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.


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Old 04-03-2008, 10:48 AM
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Default Lipoprotein preparation



Hi all,
I need a protocol for HDL and LDL preparation (if possible not using HPLC).
Thanks in advance for any help

Gianni
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Old 04-09-2008, 10:51 PM
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Default Re: Lipoprotein preparation

Hello Gianniotta,

most people use zonal or sucrose based gradient centrifugation. And collect fractions of HDL, LDL (Small dense or larger)

What exactly are you trying to do with HDL and LDL?

cheers
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Old 02-24-2014, 06:25 PM
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Default Re: Lipoprotein preparation

Selective isolation of the LDLs: mix at the vortex, 50 - 100 l of serum/plasma with 1.0 - 2.0 ml of selective precipitating reagent, amphophillic polymers, for the LDLs (dissolve in imidazole buffer 25 mM at pH 6.1, 0.4 mg/ml anionic polycyclic surfactant activator, 0.6 - 0.8 mg/ml anionic polycondensated surfactat activator and 12.4 mM polysubstituted dioxan), and incubate at 4 C for 30 min. Centrifuge at 4000 rpm (2800 g) for 5 min., discard the supernatant and pellet is resuspend in 0.5 ml of resuspending solution (0.15 M trisodium citrate and 0.11 M NaCl). For a greater purity the precipitate must be once washed with selective precipitating reagent and then centrifuge at 4000 rpms for 5 min. Suspend pellet in solubilizing solution (0.01% Triton X-100 in 100 ml of aqueous 50 g/L of NaCl) kept at 37 C, and to mix at the vortex for resuspending the LDLs. Dialyze the solution against repeated change of dialyzing solution (phosphate buffer 0.02 M at pH 7.4, 0.15 M NaCl, 0.2 g/L NaN3 and 0.1 g/L Na2EDTA). The liquid of dialysis could be also formulated as 8.4 mM K2HPO4, 1.6 mM KH2PO4, 1.34 mM EDTA and 0.65 mM of glutathione (EDTA and glutathione are added for inhibiting lipoperoxidation).
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