Thermo Fisher Scientific SARS-CoV-2 PRODUCT INFORMATION SHEET

PRODUCT INFORMATION SHEET

Intact Virus Precipitation Reagent (optimized for SARS-CoV-2)

Catalog Number 10720D

Pub. No. MAN0019857 Rev. A.0

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Product description

Biological study of viruses, including SARS-CoV-2, often requires
isolation of intact virus particles from very dilute samples. Current
isolation methods such as ultracentrifugation are tedious and
dicult. The Intact Virus Precipitation Reagent (optimized for
SARS-CoV-2) provides a simple, fast and reliable method for
concentration of intact viruses from various samples such as cell
culture media and transport media. By tying up water molecules,
this reagent forces less-soluble components (i.e., viral particles)
out of solution, allowing them to be collected after brief, low­speed centrifugation. The procedure is performed in three simple
steps and takes less than 3 hours to perform.

The provided user protocol is for SARS-CoV-2 but can be
further optimized for use with other enveloped viruses. If it
is necessary to automate the virus enrichment process, using
Dynabeads™ Intact Virus Enrichment (optimized for SARS-CoV-2)
in combination with any of the KingFisher™ instruments is
recommended.

Contents and storage

The Intact Virus Precipitation Reagent processes up to 100 mL of
cell culture media.

Components

Intact Virus Precipitation
Reagent (optimized for
SARS-CoV-2)

Amount Storage

50 mL 2–8°C

Prepare sample

1. Harvest cell culture media or virus transport media (VTM).

2. Centrifuge the media at 3,200 × g at 2–8°C for 15 minutes to

remove cells and debris.

3. Transfer the supernatant gently to a new tube without
disturbing the pellet.

Precipitate intact virus

1. Transfer the required volume of cell-free media to a new

tube and add 0.5 volumes of the Intact Virus Precipitation
Reagent (ratio 1:2 of precipitation reagent:virus-containing
media). The protocol is directly scalable and can be scaled
up or down accordingly.

Media starting volume

1 mL 500 μL

10 mL 5 mL

2. Vortex or pipette the solution up and down until there is a
homogenous solution.

3. Incubate samples at 2–8°C for 2 hours (or overnight, if
preferred).

4. Centrifuge the sample at 10,000 × g at 2–8°C for 30 minutes.

Note: Viral particles are contained in the pellet at the bottom

of the tube for swing-out rotors or on the tube wall for fixed
angle rotors (not visible in some cases).

If the intended downstream use is qPCR, reduce
centrifugation speed to 3,200 × g.

5. Aspirate and discard the supernatant.

Precipitation Reagent volume

General guidelines

• The enrichment protocol was tested on, and works reliably
for live SARS-CoV-2 virus, inactivated SARS-CoV-2 virus and
SARS-CoV-2 virus-like particles (VLP’s). Note that VLP’s do
not contain nucleic acids, and thus can only be used for
proteomic studies.

• Fetal bovine serum (FBS) contains high levels of naturally
occurring exosomes, which will be co-purified with the viral
particles due to near identical size and properties. If this is
a concern when virus is enriched from cell culture media
containing FBS, use exosome-depleted FBS in place of
normal FBS.

For Research Use Only. Not for use in diagnostic procedures.

6. Resuspend the pellet in a convenient volume of 1X PBS or
similar buer (example volumes are shown below).

Media starting volume

1 mL 25–100 μL

10 mL 100 μL to 1 mL

7. Once the pellet is resuspended, the virus is ready for
downstream analysis, ranging from functional studies to
end-point analysis of RNA and protein cargo. The enriched
viral particles can be used directly in qPCR analysis, or RNA
can be purified with MagMAX™ RNA extraction kits.

Resuspension volume

Protein analysis by electrophoresis

When performing western blot analysis, follow the standard
procedures for your electrophoresis apparatus using a wide range
protein gel (e.g., Bolt™ 4 to 12% Mini Protein Gel).

Prepare sample for electrophoresis

This is a simple protocol for sample preparation prior to
electrophoresis. Further optimization may be required for optimal
results.

1. Resuspend the enriched virus from “Precipitate intact virus”
in 30 µL of distilled water.

Note: If your antibody requires reducing conditions reduce
the volume distilled water to 26 µL, and add 4 µL 10X Bolt
Sample Reducing Agent to the sample.

2. Add 10 μL 4X Bolt™ LDS Sample Buer.

3. Heat for 10 minutes at 70°C.

4. Apply to DynaMag™ magnet for 1 minute.

5. Load supernatant containing isolated virus into the wells of

the gel for electrophoresis.

™

Related products

Product Cat. No.

Dynabeads™ Intact Virus
Enrichment (optimized for SARS-

[1]

CoV-2)

Exosome-depleted FBS A2720803

4X Bolt™ LDS Sample Buer B0007

10X Bolt™ Sample Reducing Agent B0004

Bolt™ 4 to 12%, Bis-Tris, 1.0 mm,
Mini Protein Gel, 10-well

iBlot™ 2 Gel Transfer Device IB21001

iBind™ Western System SLF1000

Goat anti-Mouse IgG1 Cross­Adsorbed Secondary Antibody,
HRP

SARS/SARS-Cov-2 Coronavirus
Nucleocapsid Monoclonal
Antibody

[1]

For use in automated protocols with KingFisher™ instruments

10700D

NW04120BOX

A10551

MA5-29981

Perform western blot

For convenience the iBlot™ 2 Gel Transfer Device can be
used for ecient blotting transfer within seven minutes after
electrophoresis without the need for liquid buers. For fast and
automated immunodetection, the iBind™ Western System can be
used.

Thermo Fisher
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The information in this guide is subject to change without notice.
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PUNITIVE, MULTIPLE, OR CONSEQUENTIAL DAMAGES IN CONNECTION WITH OR ARISING FROM THIS DOCUMENT, INCLUDING YOUR USE OF IT.
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8 January 2021