Lipids are a family of organic compounds that include monoglycerides, diglycerides, triglycerides, etc. The presence of the lipids in the biological sample is usually poorly compatible with many downstream applications, including ELISA, protease digestion of proteins, isoelectric focusing, and mass spectrometry (MS). To enable downstream analysis, it is critical to remove the lipids from the biological sample such as ascites, serum, cell & tissue culture, bile, and organ homogenates. For such purposes, traditionally, chloroform or column has been the preferred method for Delipidation. However, using the solvent removal agent or spin column to remove the lipids is tedious, timing-consuming, suffering from sample losses, environmental concerns, and challenging to adapt to high-throughput automation. We develop a novel, super-efficient lipid-specific removal agent based on magnetic beads to overcome these limitations.

BcMag™ One-Step Lipids Removal Kit uses specially designed magnetic beads to efficiently bind and remove lipids from ascites, serum, cell & tissue culture, bile, and organ homogenates. The convenient single-step, one-tube purification protocol takes only 2 minutes with very little hands-on time. The beads enable 96 samples to be processed simultaneously in less than 10 minutes.

Workflow

1.

Add the beads to a sample solution.

2.

Mix the beads with the sample by vortex or pipetting.

3.

Magnetically remove the beads.

4.

Aspirate the supernatant containing a lipid-free sample.

Features and Advantages

●

Simple protocol: No liquid transfer, One-tube, One-step

●

Ultrafast: Two-minute protocol

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Lipid-specific adsorption and high sample recovery: > 90%

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Cost-effective: Eliminates columns, centrifuge, filters, and laborious repeat pipetting

●

High-throughput: Compatible with many different automated liquid handling systems

PROTOCOL

Materials Required

Item

Magnetic Rack for centrifuge tube

** Based on sample volume, the user can choose one of the following magnetic Racks

Source

•  BcMag™ Rack-2 for holding two individual 1.5 ml centrifuge tubes (Bioclone, Cat. No. MS-01)

•  BcMag™ Rack-6 for holding six individual 1.5 ml centrifuge tubes (Bioclone, Cat. No. MS-02)

•  BcMag™ Rack-24 for holding twenty-four individual 1.5-2.0 ml centrifuge tubes (Bioclone, Cat. No. MS-03)

•  BcMag™ Rack-50 for holding one 50 ml centrifuge tube, one 15 ml centrifuge tube, and four individual 1.5 ml centrifuge tubes (Bioclone, Cat. No. MS-04)

Item

BcMag™ 96-well Plate Magnetic Rack.

Source

•  BcMa™ 96-well Plate Magnetic Rack (side-pull) compatible with 96-well PCR plate and 96-well microplate or other compatible Racks (Bioclone, Cat. No. MS-05)

Item

Adjustable Single and Multichannel Pipettes

Item

Centrifuge with Swinging Bucket

Addition items are required if using 96-well PCR plates / tubes

Vortex Mixer

** The user can also use other compatible vortex mixers. However, the Time and speed should be optimized, and the mixer should be: Orbit ≥1.5 mm-4 mm, Speed ≥ 2000 rpm

Eppendorf™ MixMate™

Eppendorf, Cat. No. 5353000529

Tube Holder PCR 96

Eppendorf, Cat. No. 022674005

Tube Holder 1.5/2.0 mL, for 24 × 1.5 mL or 2.0 mL

Eppendorf, Cat. No. 022674048

Smart Mixer, Multi Shaker

BenchTop Lab Systems, Cat. No. 5353000529

1.5/2.0 mL centrifuge tube

96-well PCR Plates or 8-Strip PCR Tubes

PCR plates/tubes

** IMPORTANT! If using other tubes or PCR plates, make sure that the well diameter at the bottom of the conical section of PCR Tubes or PCR plates must be ≥2.5mm.

Items

Magnetic Rack for centrifuge tube

** Based on sample volume, the user can choose one of the following magnetic Racks

Source

●

BcMag™ Rack-2 for holding two individual 1.5 ml centrifuge tubes (Bioclone, Cat. No. MS-01)

●

BcMag™ Rack-6 for holding six individual 1.5 ml centrifuge tubes (Bioclone, Cat. No. MS-02)

●

BcMag™ Rack-24 for holding twenty-four individual 1.5-2.0 ml centrifuge tubes (Bioclone, Cat. No. MS-03)

●

BcMag™ Rack-50 for holding one 50 ml centrifuge tube, one 15 ml centrifuge tube, and four individual 1.5 ml centrifuge tubes (Bioclone, Cat. No. MS-04)

BcMag™ 96-well Plate Magnetic Rack

●

BcMa™ 96-well Plate Magnetic Rack (side-pull) compatible with 96-well PCR plate and 96-well microplate or other compatible Racks (Bioclone, Cat. No. MS-05)

Adjustable Single and Multichannel Pipettes

Centrifuge with Swinging Bucket

Addition items are required if using 96-well PCR plates/tubes

Vortex Mixer

** The user can also use other compatible vortex mixers. However, the Time and Speed should be optimized, and the mixer should be: Orbit ≥1.5 mm-4 mm, Speed ≥ 2000 rpm

Eppendorf™ MixMate™

Tube Holder PCR 96

Tube Holder 1.5/2.0 mL, for 24 × 1.5 mL or 2.0 mL

Smart Mixer, Multi Shaker

Eppendorf, Cat. No. 5353000529

Eppendorf, Cat. No. 022674005

Eppendorf, Cat. No. 022674048

BenchTop Lab Systems, Cat. No. 5353000529

Eppendorf™ MixMate™

Tube Holder PCR 96

Tube Holder 1.5/2.0 mL, for 24 × 1.5 mL or 2.0 mL

Eppendorf, Cat. No. 5353000529

Eppendorf, Cat. No. 022674005

BenchTop Lab Systems, Cat. No. 5353000529

1.5/2.0 mL centrifuge tube

96-well PCR Plates or 8-Strip PCR Tubes

PCR Plates/Tubes

! IMPORTANT ! If using other tubes or PCR plates, ensure that the well diameter at the bottom of the conical section of PCR Tubes or PCR plates must be ≥2.5mm.

The following protocol is an example. The beads and sample volume can be rational Scale-up (or down). Do not use buffers containing organic solvents.

Procedure

1.

Shake the bottle to resuspend the magnetic beads until it is homogeneous entirely.

2.

Add 10μl magnetic beads to a 100 μl sample containing lipids.

3.

Mix the sample with beads for 1-2 minutes by slowly pipetting up and down 20-25 times or vortex for 5 minutes at 2000 rpm for PCR plates or 800 rpm for Elisa plates.

4.

Place the sample plate or tube on the magnetic separation plate for 30 seconds or until the solution is clear.

5.

Transfer the supernatant to a clean plate/tube while the sample plate remains on the magnetic separation plate. The sample is ready for downstream applications.

Learn More

Instruction Manual

MSDS