AB Vector - GFP reporter

  GFP reporter
 
  GFP reporter (Aequorea victoria green-fluorescent protein) (Acc# AAA58246) is encoded in majority of our linearized baculovirus DNA vectors and recombinant baculoviruses. Their list is provided below. GFP allows monitoring of virus propagation, optimization of cultivation conditions, titration of virus stocks by "terminal green dilution", purification of GST-tagged proteins using GFP-Trap affinity reagents. For your convenience, we provide Green Control, which is a titrated reference stock of a baculovirus expressing GFP. GFP saves time and effort; there is no need for expensive baculovirus titration kits or specialized equipment. The only essential pieces required are the same 365 nm UV transilluminator and digital camera routinely used for ethidium bromide stained gels. Alternatively, you can determine virus titers by counting green foci of infected cells using fluorescent microscopy.

In addition, you can visualize GFP expressing cells by electron or light microscopy (Luby-Phelps et al., J. Histochem Cytochem. 51: 271-274, 2003), titer GFP-expressing vectors using flow cytometry (Hitt et al., Mol. Biotechnol., 14: 197-203, 2000), or use GFP to study the spread of nucleopolyhedrovirus infection in insects (Barrett et al., Tissue Cell, 30: 602-616, 1998).
 
 
 
  Our pVL-GFP™ vector allows expression of target proteins in-fusion with GFP tag. The GFP tag is widely used for visualization and isolation of protein complexes, studies of protein-protein interactions, protein trafficking within the cells, and mechanisms of protein degradation (Cristea et al., Mol Cell Proteomics, 4:1933-41, 2005; Michaelson and Philips, Methods Enzymol, 406:296-315, 2006; Hayes et al., Cancer Lett., 206:129-35, 2004 ; Takeuchi et al., EMBO J., 26:123-31, 2007).

Especially useful is the ability of the GFP tag to misfold and loose its fluorescence if its fusion partner is misfolded. This property was used for selection of target protein mutants with improved solubility (Van den Berg et at, J Biotechnol, 121:291-8, 2006), for finding soluble proteins derived from randomly fragmented cDNAs (Nakayama and Ohara, Biochem Biophys Res Commun, 312:825-30, 2003), and in studies of protein aggregation (Schrodel and de Marco, BMC Biochem, 6:10, 2005).

Of particular interest are studies of GFP fusion aggregates with proteins related to protein folding diseases (Krobitsch and Lindquist, Proc Natl Acad Sci U S A, 97:1589-94, 2000; Wang et al., Hum Mol Genet., 14:3673-84, 2005; Pandey et al., Exp Neurol., 197:515-20, 2006; Jiang et al., J Biol Chem., 282:7912-20, 2007). Such fusions could be used for high throughput screening of compounds that inhibit aggregation of these important drug targets (Gazit, ASC Chemical Biology, 1:417-19, 2006). AB Vector offers some protein folding diseases targets in-fusion with GFP.
 
  Products equipped with GFP reporter
 
Kits                                                                         Click on the product name for technical information!
Cat # Product Specification Unit size Price*
K20 Green Kit Great value, all the vectors and a control you need to start
High level target protein expression controlled by polyhedrin promoter
GFP marker to simplify manipulations
Compatible with Sf9, Sf21, Hi-Five insect cells and with wide range of      transfection reagents and media recommended for baculovirus work
     Includes:

     1. ProGreen™ GFP+ baculovirus vector DNA, 25 μl, 2.5 μg
     2. pVL1393 plasmid transfer vector, 50 μl, 25 μg
     3. GC (green control) GFP+ control recombinant baculovirus
NA
  $490.00
K22 C2 Kit Recommended for expression of difficult cytoplasmic proteins and proteins
     with large cytoplasmic domains
     Includes:

     1. ProFold™-C2 baculovirus vector DNA, 25 μl, 2.5 μg
     2. pVL1393 plasmid transfer vector, 50 μl, 25 μg
NA $895.00
K23 ER1 Kit Recommended for expression of glycoproteins using native signal sequences
     Includes:

     1. ProFold™-ER1 baculovirus vector DNA, 5 transfections
     2. pVL1393 plasmid transfer vector, 50 μl, 25 μg
NA $895.00
K24 ER1-bee Kit Recommended for expression of glycoproteins using powerful honeybee
     melittin signal sequence
     Includes:

     1. ProFold™-ER1 baculovirus vector DNA, 5 transfections
     2. pAcAB3 plasmid transfer vector, 50 μl, 25 μg
NA $995.00
 
Plasmid transfer vectors                               Click on the product name for technical information!
Cat # Product Specification Unit size Price*
B8 pVL-GFP™ High level expression of proteins with C-terminal GFP tag 50 μl, 0.5 μg/μl $395.00
 
Linearized baculovirus vector DNA
CsCl purified and digested with Bsu 36.I                  Click on the product name for technical information!
Cat # Product Specification Unit size Price*
A1 ProGreen™ High level of target protein expression
GFP marker to monitor virus propagation
25 μl, 2.5 μg
5 transfections
$295.00
A3 ProFold™-C2 Overexpressed Hsp40 and Hsc70 chaperones facilitate target protein folding in the      cytoplasm
GFP marker to monitor virus propagation
Slightly reduced level of protein expression
25 μl, 2.5 μg
5 transfections
$695.00
A4 ProFold™-ER1 Overexpressed calreticulin facilitates glycoprotein folding in the ER
Overexpressed PDI facilitates folding of cysteine-rich proteins
GFP marker to monitor virus propagation
High level of target protein expression
25 μl, 2.5 μg
5 transfections
$695.00
 
Controls                                                                    Click on the product name for technical information!
Cat # Product Specification Unit size Price*
C14 GC (green control) Recombinant baculovirus stock Positive control baculovirus in experiments where GFP fluorescence is used to      monitor virus propagation, optimize cultivation conditions, or titrate of virus stocks 1 ml, 108 p.f.u./ml $195.00
C15 GCP (green control plasmid) Plasmid transfer vector DNA Positive control for transfection with linearized GFP- baculovirus DNA. Green      fluorescence of cells is indicative of successful transfection.
50 μl, 0.2 μg/μl $145.00
 
Recombinant baculovirus stocks for expression of molecular chaperones
Cat # Product Specification Unit size Price*
H6 FoldHelper™-ER3 Overexpressed calreticulin to facilitate glycoprotein folding in the ER
Overexpressed PDI to facilitate folding of cysteine-rich proteins
Overexpressed PPI to facilitate isomerization of imide bonds in proline-rich
     proteins
GFP marker to monitor virus propagation
2 ml,5x107p.f.u./ml $695.00
 
Recombinant baculovirus stocks for expression of protein folding diseases targets in-fusion with GFP
Cat # Product Specification Unit size Price*
PD21 BV-Syn-GFP
Recombinant baculovirus stock
Expression of alpha synuclein-GFP fusion protein 1 ml, 108 p.f.u./ml $899.00
PD29 BV-SOD1-GFP
Recombinant baculovirus stock
Expression of SOD1-GFP fusion protein 1 ml, 108 p.f.u./ml $899.00
 
  *Go to PRODUCTS to order online. Alternatively, email an order as pdf attachment to info@abvector.com, or send it by Fax to 1-760-454-2465 or call 1-866-683-2867 to order by phone.
 
  If you have not worked with recombinant baculoviruses, you can get a quick update at TECHNOLOGY. Go to BACULOVIRUS TUTORIAL for simple on-line instructions on baculovirus transfection, propagation of recombinant baculoviruses and recombinant baculovirus protein expression studies.
 
  Click on PRODUCTS to view the entire list of recombinant baculovirus-related products.
  Commercial use of GFP reporter requires licensing.


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