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Adenoviral Titration Kits

 
 

Clontech offers two kits for rapidly titrating recombinant adenovirus:

  • The Adeno-X™ Rapid Titer Kit allows functional, hexon-based titration
  • The Adeno-X qPCR Titration Kit allows ultra-fast, qPCR-based titration

Adeno-X™ Rapid Titer Kit

The Adeno-X Rapid Titer Kit provides a quick and simple antibody based-assay that can be used to titrate any first-generation E1-deleted adenovirus (1). The 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 (2) 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 (Figure 1).

Adeno-X qPCR Titration Kit

The Adeno-X qPCR Titration Kit provides an extremely 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 SYBR® 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 (Figure 2). 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.

 
 
 
 

Figure 1. Adeno-X Rapid Titer Kit protocol.

 
 

Figure 2. Flowchart of the procedures used for titrating adenoviral DNA with the Adeno-X qPCR Titration Kit.

 
 
 
 

Applications

  • Titrate any adenoviral stock 
  • References

    1. Bewig, B. & Schmit, W. E., et al. (2000) BioTechniques 28:871–873.
    2. Graham, F. L., et al. (1977) J. Gen. Virol. 36:59–72.

    Components

    Adeno-X Rapid Titer Kits (Cat. No. 632250)
    Mouse Anti-Hexon Antibody
    Rat Anti-Mouse Antibody (HRP Conjugate)
    DAB Substrate and Buffer

    Adeno-X qPCR Titration Kit (Cat. No. 632252)
    • Adeno-X qPCR Titration Components (not sold separately)
      • Adeno-X DNA Control Template
      • Adeno-X Forward Titer Primer
      • Adeno-X Reverse Titer Primer
      • DNase I
      • EASY Dilution Buffer
    • SYBR® Advantage® qPCR Premix (Cat. No. 639676)
      • 2X SYBR® Advantage® qPCR Premix
      • 50X ROX Reference Dye LSR
      • 50X ROX Reference Dye LMP
    • NucleoSpin® Virus Kit (Cat. No. 740977.10)
      • Lysis Buffer RAV1
      • Wash Buffer RAW
      • Wash Buffer RAV3
      • RNase-free H2O
      • Elution Buffer RE
      • Carrier RNA
      • Proteinase K
      • Proteinase Buffer PB
      • NucleoSpin® Virus Columns
      • Collection Tubes

    Storage Conditions

    Adeno-X Rapid Titer Kit
    −20oC for Mouse Anti-hexon Antibody and 10X DAB Substrate
    4oC for Rat Anti-Mouse Antibody (HRP Conjugate) and Stable Peroxidase Buffer

    Adeno-X qPCR Titration Kit
    For Adeno-X Titration Components
    −20oC for all components

    For SYBR Advantage qPCR Premix
    −70oC in a dark container
    After thawing, store at 4oC in a dark container—do not refreeze!

    For NucleoSpin Virus Kit
    Kit components are stable at room temperature for 1 year.
    See the User Manual for storage of prepared buffers

     
     
     
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    Adeno-X Rapid Titer Kit 120 titrations 632250 $375.00 USD    Add to cart
     
    Adeno-X qPCR Titration Kit 200 rxns 632252 $396.00 USD    Add to cart
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