AB Vector - Cytoplasmic proteins

  Cytoplasmic proteins
 
  Overexpressed cytoplasmic proteins are often misfolded which results in the accumulation of biologically inactive aggregates (inclusion bodies). In unmodified cells these proteins do not aggregate since they are expressed at low yield (typically below 0.1% of the total cell protein). At such low concentrations molecular chaperones are in excess to the newly synthesized proteins which ensures their correct folding. However, if a protein is overexpressed, its concentration by far exceeds concentration of molecular chaperones in the cytoplasm, resulting in target protein misfolding and aggregation.
ProFold™-C1 and ProFold™-C2 linearized baculovirus vector DNA can provide for overexpression of both a cytoplasmic protein of interest and major cytoplasmic molecular chaperones Hsp40 and Hsc70. This often improves the yield of correctly folded cytoplasmic target proteins as compared to the same target proteins expressed using conventional vectors (Bac-to-Bac™**, BaculoGold™***, and BacPAK6™****) which do not provide molecular chaperones. Click on TECHNOLOGY for the details.
 
  Mix and match features provided in the vectors
 

Use pVL1393 plasmid transfer vector if you wish to express target proteins without affinity tags.

Use pAB-6xHis™ plasmid transfer vector if you wish to express target proteins with a polyhistidine tag.


Use pAB-GST™ plasmid transfer vector if you wish to express target proteins with a GST tag.


Use pAB-MBP™ plasmid transfer vector if you wish to express target proteins with a maltose binding protein (MBP) tag. Note that an MBP tag could improve protein solubility.


Combine any of the above plasmid transfer vectors with ProFold™-C1 linearized baculovirus DNA if you wish to provide Hsp40 and Hsc70 molecular chaperones.
  The above features can be mixed and matched in various combinations, while supporting a high level of target protein expression. They can also be combined with "plain vectors" which only provide a high level of protein expression (pVL1393, ProEasy™, ProGreen™, ProFold™-0). A plasmid transfer vector is always combined with a baculovirus DNA vector, and some combinations are available as Kits. Click on TECHNOLOGY for more information on baculovirus expression system. Go to BACULOVIRUS TUTORIAL for simple on-line instructions on baculovirus transfection, propagation of recombinant baculoviruses and recombinant baculovirus protein expression studies for details.
 
  Examples
 
  1.Solubility of a cytoplasmic protein is a problem. Insert gene encoding target cytoplasmic protein in pVL1393 plasmid vector if no affinity tag is required. Alternatively, use the pAB-GST™, pAB-6xHis™, or pAB-MBP™ vectors to provide target proteins with GST, His, or MBP tags. Co-transfect plasmid construct with ProFold™-C12 linearized baculovirus DNA.
  2.Target cytoplasmic protein is not problematic. Use ProEasy™ linearized baculovirus DNA, as it is the easiest method to construct recombinant baculoviruses. It is compatible with any of the pVL1393, pAB-GST™, pAB-6xHis™, or pAB-MBP™ plasmid transfer vectors.
  3. You have little experience with recombinant baculoviruses. Get a quick update on relevant recombinant baculovirus technologies at Materials and methods. Use ProEasy™ linearized baculovirus DNA, as it is the easiest method to construct recombinant baculoviruses. It is compatible with any of the pVL1393, pAB-GST™, pAB-6xHis™, or pAB-MBP™ plasmid transfer vectors.
 
  Relevant products
 
Plasmid transfer vectors                             Click on the product name for technical information!
Cat # Product Specification Unit size Price*
B1 pVL1393 High level of target protein expression 50 μl, 0.5 μg/μl $295.00
B4 pAB-6xHis™ High level expression of proteins with cleavable N-terminal 6xHis tag 50 μl, 0.5 μg/μl $295.00
B5 pAB-GST™ High level expression of proteins with cleavable N-terminal GST tag 50 μl, 0.5 μg/μl $295.00
B6 pAB-MBP™ High level expression of proteins with cleavable N-terminal MBP tag
MBP tag improves protein solubility and yield
50 μl, 0.5 μg/μl $395.00
B7 pAB-6xHis-MBP™ High level expression of proteins with cleavable N-terminal 6xHis MBP tag
High level expression of proteins with C-terminal MBP tag
MBP tag improves protein solubility and yield
50 μl, 0.5 μg/μl $495.00
 
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™-C1 Overexpressed Hsp40 and Hsc70 chaperones facilitate target protein folding in the      cytoplasm
GFP marker to monitor virus propagation
High level of target protein expression
25 μl, 2.5 μg
5 transfections
$695.00
A9 ProFold™-0 The best value baculovirus vector
High level of target protein expression
25 μl, 2.5 μg
5 transfections
$195.00
A10S ProEasy™ The easiest generation of recombinant baculoviruses
Ideal for HTP applications
High level of target protein expression
25 μl, 2.5 μg
5 transfections
$599.00
 
Controls                                                                    Click on the product name for technical information!
Cat # Product Specification Unit size Price*
C13 NC (negative control) Recombinant baculovirus stock Extracts of cells infected with NC is an ideal negative control as there is no      polyhedrin or target protein expression
NC extracts are run side by side in SDS-PAGE with extracts of cells infected with      recombinant baculoviruses
Recombinant protein is detected as a prominent band which is absent in NC      extracts
1 ml, 108 p.f.u./ml $195.00
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 $295.00
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
 
  *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 baculovirus expression system, also referred to as baculovirus expression vector system or BEVS, 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.


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