Takara BacPAK™ p35 ELISA Kit (Single Wash), 96 Rxns

BacPAK p35 ELISA Kit (Single Wash) enables you to determine functional baculovirus titers based on the expression of baculoviral p35. The ELISA protocol has been developed to reduce the number of steps, which minimizes hands-on time. The entire protocol can be completed in 20 hours, including an overnight infection step. However, only 2 hrs of hands-on time is required. The resulting p35 titer value can easily be converted to IFU/mL with ourBaculovirus p35-to-IFU calculator.

Cat. # 631477 contains an Anti-p35 Coated Plate of 96 wells (available as 12 x 8-well strips), p35 Control, Lysis Buffer, p35 Buffer, Anti-p35 (HRP Conjugate), Washing Buffer (20X), TMB Substrate, and Stop Solution.

Comparison of baculovirus titration methods.A strong correlation was observed between the physical titration methods (GoStix and qPCR) and the functional methods that require infection (ELISA and plaque staining). The time required to obtain results for each method is summarized in the table.

ZsGreen1-expressing baculovirus was generated using the BacPAK Baculovirus Expression System (Cat. # 631402), following the manufacturer’s protocol. Baculovirus supernatant was harvested from several plaques and amplified in fresh Sf21 cells (P1) for four days.

The baculovirus supernatant from a single clone was analyzed using two functional titration methods, the BacPAK Baculovirus Rapid Titer Kit (Cat. # 631406) and the BacPAK p35 ELISA Kit (Single Wash) (Cat. # 631477), as well as two physical titration methods, the BacPAK qPCR Titration Kit (Cat. # 631414) and the BacPAK Baculovirus GoStix Plus (Cat. # 631479).

Infectious units (IFU) and GoStix results from Clone 11 (data not shown) were used as a reference with our online calculator to convert the units from the different methods into IFU for Clone 4 (shown here). The formula used for these conversions is available in the user manual.

Timelines associated with most commonly used methods of baculoviral vector titration.BacPAK GoStix Plus and ELISA methods are significantly faster than traditional methods and DO not require the special equipment of qPCR- and FACS-based methods. FACS = fluorescence-activated cell sorting.

BacPAK GoStix Plus and ELISA baculovirus titration methods can be used at various stages in your baculovirus workflow.BacPAK Baculovirus GoStix Plus (A) can be used for quick titer checks in stages such as clone screening or starting from a stored viral stock for downstream production. BacPAK p35 ELISA Kit (Single Wash) (B) can be used to determine functional titers in steps such as baculovirus titer normalization and expression optimization. You can also use these two products in other stages of your workflow.