FAQ - Adenovirus Production

Here are our most-asked questions about virus handling and our protocol!


Virus Use
and Handling

How do I thaw and aliquot my samples?

We supply the virus in 1 mL aliquots and at 1e12 particles/mL. We recommend thawing the virus at room temperature and then aliquoting at least 50 uL in 0.5 mL screw cap tubes. Do not make aliquots in PCR tubes or eppendorf centrifuge tubes. Do not store or thaw the virus on ice, -20°C or 4°C. We do offer custom aliquoting at the time of manufacture – please inquire.

What buffer is my sample in?

Our standard buffer is A195 buffer: 10 mM Tris, 0.1 mM EDTA, 1 mM MgCl2, 10 mM Histidine, 75 mM NaCl, 5% sucrose, 0.02% PS-80, 0.50% EtOH, pH 7.4. However, we are happy to dialyze into the buffer of your choice!

Can I dilute the sample before injecting mice?

Yes. Common diluents include PBS and TBS. Once you dilute your sample, do NOT refreeze! The diluted virus means you have also diluted the cryoprotectants. A freeze/thaw cycle with diluted cryoprotectant will result in dead virus on the next thaw. Also, do not store or thaw the virus on ice, 4°C, or -20°C. -80° C or room temperature is best.

Can I use my virus both for both in-vivo and in-vitro experiments?

Yes! The virus you receive from us will be purified over 2 rounds of CsCl gradients and formulated at 1e12 particles per mL in A195 buffer. We guarantee at least 1e10 pfu/mL. It is perfectly suited for either in-vivo or in-vitro use without any additional modification (but dilution in the case of in-vitro experiments.)

Can I use my virus to propegate more virus?

Yes, for your laboratory use. We ask that you not distribute the virus to other laboratories or institutions.

Virus Ordering and Shipping

How long will it take to produce my adenovirus?

Amplification of existing particles typically takes ~10 days. If we start with shuttle plasmids turnaround time is generally 3 to 4 weeks.

What cells are used to grow my virus?

We almost always use HEK 293 cells. Any virus that is E1 deleted must be grown in HEK cells. However, A549 and Hela cells are used when a virus is replication competent (as in WT) or conditionally competent (replicates in tumor cells, A549s). For viruses containing modified E1a and E1b, we amplify in A549 cells to prevent possible recombination with E1 sequences in HEK cells.

If I run out of virus, how long will it take to get more?

We try and always keep a stock of your purified particles in our freezers, so generally we can ship an aliquot of purified particles to you the next day. If we do not to have your materials in stock, we can generally amplify and purify a new prep for you in ~10 days.

What is the miniumum order volume for virus particles?

1 mL

Where do I find a list of premade viruses on your site?

Viruses that are widely used are available off the shelf, CsCl purified, contain 1e12 particles/mL and have been tested for RCA and PFU titered. These virus particles can be found on our products page. Most of the viruses we make are custom constructs for our individual clients and these materials are held in confidence. These viruses may be available with permission from the originating PI. Please inquire for specific genes and we will determine how we can help.

How do I ship my samples to Antibodies Inc?

All virus samples should be sent via overnight carrier on dry ice. Please email the tracking number to info@antibodiesinc.com. DNA samples can be sent overnight in water, 10 mM TRIS, or TE on cold packs. Please see our contact page for complete shipping address.

Virus Data

Will I receive my PFU titers?

PFU (plaque-forming units) titers are lot-specific and take about 10 days to perform these assays. PFU titers and the final characterizaton forms are emailed when these assays are complete and characterization forms available for every lot number.

Why is my particles number different from my PFU?

The particles number is the physical titer based on genome content, measuring DNA and genomes. Not all virus that has genomes is infectious, and we have a plaque assay that measures infectious titer, or PFU (plaque forming units). Particles are always higher than PFU, and ratios can vary. For example, a typical particle:PFU ratio for GFP is 30:1, but this ratio isn't the same for all samples.

Should I use particle number or PFU in my experiments?

You will find both instances in the literature - some groups use only paricle titer and others use PFU titer. The important thing is to use one or the other.

What if I need the amino acid sequence?

We are happy to provide sequences upon request.

Have a different question? E-mail us at: orders@antibodiesinc.com

Ready to get started? Contact us for a quote today!

Request a quote here.

Read additional FAQs here.

See an overview of our adenovirus production service here.

Ready to Get Started?

For questions or a quote from one of our scientists, submit your project details below. We will respond within two business days. If you require immediate assistance, call us at (800) 824-8540.