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Selection guides Virus kits: product finder tool

Adenovirus titration kits

Adenovirus titration kits

  • The Adeno-X Rapid Titer Kit allows functional, hexon-based adenovirus titration.
  • The Adeno-X qPCR Titration Kit allows ultra-fast adenovirus titration using quantitative PCR.
  • Each Adeno-X GoStix cassette is a sensitive instant test to detect adenovirus early during amplification and to determine the optimum harvest time.

  • The Adeno-X Rapid Titer Kit allows functional, hexon-based adenovirus titration
  • The Adeno-X qPCR Titration Kit allows ultra-fast adenovirus titration using quantitative PCR
  • Each Adeno-X GoStix cassette is a sensitive instant test to detect adenovirus early during amplification and to determine the optimum harvest time

Adeno-X Rapid Titer Kit

This kit is designed around a hexon-specific antibody, which is used to label infected cells. Hexon protein is encoded by the adenoviral genome and is an essential component of the adenoviral capsid required for adenoviral replication, but its expression depends on the E1 gene product. Thus, E1 trans-complementing cell types such as HEK 293 can be used to measure infectious activity because only infected cells will produce the hexon protein. The rapid titer assay takes 1–2 hours to set up and another 3 hours (two days later) to label, stain, and count infected cells. Each stained cell corresponds to a single infectious unit.

Adeno-X qPCR Titration Kit

The Adeno-X qPCR Titration Kit provides a fast, simple, and accurate method for titrating adenoviral stocks from all Ad5-based adenoviral vectors, including recombinant adenovirus created with our Adeno-X expression systems. The protocol combines qPCR and TB Green chemistry, allowing you to determine the viral genome copy number in adenoviral preparations (i.e., crude lysates or purified stocks) from a calibrated DNA standard curve. The kit delivers results in just 4 hours, and because qPCR titration is so fast, target cells can be infected with accurately titrated virus on the same day the virus is harvested.

Adeno-X GoStix

Adeno-X GoStix will rapidly (~10 min) tell you if you are ready to go with your adenovirus prep and is ideal for determining the best time to harvest your virus following amplification. Simply apply 20 µl of culture supernatant to the GoStix cassette, chase it through with four drops of buffer and wait 2–20 min for a band to appear in the window of the cassette. The amount of adenovirus detected in the supernatant correlates strongly with adenovirus production inside the cells. Each test is sensitive enough to detect adenovirus in a small amount of crude supernatant and is also used to ensure maximum yields during the adenovirus amplification process (we employ these same test cassettes to determine the optimum time to harvest virus from infected cells). Just 20 µl of supernatant is tested each day following infection of HEK 293 cells. The presence of a strong band in the cassette window ensures that high-titer adenovirus is harvested at the peak time.

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Cat. # Product Size Price License Quantity Details
632270 Adeno-X™ GoStix™ 20 Tests USD $390.00

License Statement

ID Number  
89 This product is protected by U.S. Patent 9,945,850 and additional pending patent. For further license information, please contact a Takara Bio USA licensing representative by email at licensing@takarabio.com.

Adeno-X GoStix are designed to instantly confirm the presence of adenovirus in cell culture supernatants or lysates. The test detects adenoviral hexon proteins and can be used to determine if infectious virus was produced post-transfection, or for selecting the best time to harvest your virus. The included Hexon Control provides confirmation of GoStix test function.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Documents Components You May Also Like Image Data Resources

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Early detection of adenovirus during rescue

Early detection of adenovirus during rescue

Early detection of adenovirus during rescue. Panel A. The Adeno-X GoStix protocol takes only a couple of minutes to indicate the presence of adenovirus particles. Supernatant from transfected cells is placed on the cassette, followed by chase buffer. Panel B. Adeno-X GoStix were used to confirm virus production during the rescue procedure. PacI-digested adenovirus expressing ZsGreen1 was transfected into HEK 293 cells, which were monitored for the emergence of cytopathic effects by phase and fluorescence microscopy (middle row). 20 μl of culture medium was applied to the Adeno-X GoStix on each day (bottom row). Four days post-transfection, Adeno-X GoStix were able to detect virus in culture supernatants containing plaques that were only weakly perceptible under phase microscopy and confirmed by fluorescence microscopy.

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Rapid adenovirus detection during amplification

Rapid adenovirus detection during amplification

Rapid adenovirus detection during amplification. Panel A. Adenovirus expressing ZsGreen1 was amplified in HEK293 cells. 48 hr postinfection, and each day thereafter, the supernatant and the cell pellet were harvested, and titers were quantified by counting fluorescent cells in an infectious unit (IFU) assay. The increase in virus level in the cell pellets was mirrored by a proportional increase in the amount of virus collected from the culture supernatant. The amount of virus in the supernatant remained < 10% of the total virus output. Panel B. Supernatant serves as a clear indicator of total yields. In addition to IFU determination, 20 μl of each sample was applied to an Adeno-X GoStix. The increase in band intensity paralleled the increase in virus production observed by infectious unit titer.

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Use Adeno-X GoStix to instantly confirm the presence of adenovirus in cell culture supernatants or lysates

Use Adeno-X GoStix to instantly confirm the presence of adenovirus in cell culture supernatants or lysates
Use Adeno-X GoStix to instantly confirm the presence of adenovirus in cell culture supernatants or lysates. The test, which detects adenoviral hexon proteins, requires just 20 µl of adenoviral supernatant to determine if infectious virus was produced posttransfection, or to select the best time to harvest your virus following amplification. The included Hexon Control (bottom) confirms GoStix test function.

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Constructing recombinant adenovirus with In-Fusion Cloning technology

Constructing recombinant adenovirus with In-Fusion Cloning technology

Constructing recombinant adenovirus with In-Fusion Cloning technology. DNA sequences can be rapidly transferred as PCR products to any pAdenoX vector using the In-Fusion cloning method. In this example, your gene of interest is amplified with 15 bp extensions that are homologous to the ends of the linearized adenoviral vector. The PCR product is then purified and mixed with the linearized adenoviral vector of choice in the In-Fusion reaction. Following the reaction, a portion of the mixture is transformed into E. coli (Stellar Competent Cells) and screened. Once a PCR-positive clone is identified, the recombinant pAdenoX vector is amplified, purified, and subsequently linearized with the restriction enzyme PacI, then transfected into Adeno-X 293 cells for viral rescue and amplification. Adeno-X GoStix can be used to determine the status of adenovirus rescue.

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What to expect when measuring viral titer with Lenti-X or Adeno-X GoStix

What to expect when measuring viral titer with Lenti-X or Adeno-X GoStix

What to expect when measuring viral titer with Lenti-X or Adeno-X GoStix. The control band (C) will always be detected, the test band shows (T) positive only you if there is enough virus in your sample.

632252 Adeno-X™ qPCR Titration Kit Each USD $490.00

The Adeno-X qPCR Titration Kit is designed to be used with real-time PCR instruments to rapidly titrate adenoviral stocks from all Ad5-based adenoviral vectors. The protocol combines qPCR with green intercalating dye chemistry, allowing you to determine the viral genome copy number in adenoviral preparations (i.e., crude lysates or purified stocks) from a calibrated DNA standard curve. The kit includes a viral nucleic acid purification kit, a calibrated Adeno-X DNA Control Template, primers, and all the reagents required to conduct 200 TB Green-based qPCR reactions with either the DNA Control Template or purified viral DNA.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Documents Components Image Data Resources

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Flowchart of the procedures used for titrating adenoviral DNA with the Adeno-X qPCR Titration Kit

Flowchart of the procedures used for titrating adenoviral DNA with the Adeno-X qPCR Titration Kit
Flowchart of the procedures used for titrating adenoviral DNA with the Adeno-X qPCR Titration Kit.

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Adeno-X Rapid Titer Kit protocol

Adeno-X Rapid Titer Kit protocol
Adeno-X Rapid Titer Kit protocol.
632250 Adeno-X™ Rapid Titer Kit 120 Titrations USD $525.00

The Adeno-X Rapid Titer Kit takes advantage of production of viral hexon proteins for the titration of adenoviral stocks in 48 hours. A viral stock is tested by making serial dilutions, infecting HEK 293 cells, and 48 hours later, fixing and staining those cells with the Mouse Anti-Hexon Antibody. The signal is detected with a Rat Anti-Mouse Antibody conjugated to horseradish peroxidase (HRP) and developed with metal-enhanced 3,3'- Diaminobenzidine tetrahydrochloride (DAB).

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Documents Components Image Data Resources

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Flowchart of the procedures used for titrating adenoviral DNA with the Adeno-X qPCR Titration Kit

Flowchart of the procedures used for titrating adenoviral DNA with the Adeno-X qPCR Titration Kit
Flowchart of the procedures used for titrating adenoviral DNA with the Adeno-X qPCR Titration Kit.

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Adeno-X Rapid Titer Kit protocol

Adeno-X Rapid Titer Kit protocol
Adeno-X Rapid Titer Kit protocol.

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632250: Adeno-X Rapid Titer Kit

632250: Adeno-X Rapid Titer Kit

Technical notes Adenoviral titration by qPCR
Increase the concentration of adenovirus receptor CAR on your target cells Adenovirus receptor booster
Adenoviral vectors and systems Adenoviral vector systems

Overview

  • Obtain a functional titer in 4 hours using either the Adeno-X Rapid Titer Kit (hexon detection) or Adeno-X qPCR Titration Kit (genome detection)
  • More precise and less time-consuming than a plaque or end-point dilution assay
  • Obtain a qualitative measurement of adenovirus in 10 minutes using Adeno-X GoStix
  • Use Adeno-X GoStix to quickly determine the optimal harvest time

More Information

Please see the product's Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.


Accurately determine adenovirus titer using qPCR

Consistent data requires consistent titer

The Adeno-X qPCR Titration Kit combines qPCR with TB Green chemistry, allowing you to accurately determine the viral genome copy number in adenoviral preparations from a calibrated DNA standard curve.

See the data Viral transduction learning center

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