AB Vector - Inclusion bodies problem

  Inclusion bodies problem
 
  Inclusion bodies are a common outcome of protein overexpression. Overexpressed proteins are prone to aggregation as the concentration of molecular chaperones in the host cells is not sufficient to shield partially folded hydrophobic areas of nascent target proteins from interacting. Incorrect formation of disulfide bonds may also contribute to the problem. Unlike conventional E.coli or baculovirus vectors, ProFold™-C2, ProFold™-ER1, ProFold™-PDI vectors facilitate protein folding due to co-expression of large amounts of certain chaperones alongside with the overexpressed protein of interest.
 
  An additional method of reducing protein aggregation is to provide the protein with a maltose binding protein (MBP) tag. MBP is a highly soluble protein which is known to improve solubility of MBP-target protein fusions (Lee et al., 2006, Protein Pept Lett. 13(5):431-5, 2006; Korepanova et al., Protein Expr Purif. 53(1):24-30, 2007). The MBP tag can be provided on the amino- terminus of a target protein using our pAB-MBP™ vector or either the amino- or carboxy- terminus using our pAB-6xHis-MBP™ vector. The fusion protein can be isolated using affinity purification on amylose or immobilized metal resin, and the tag can be cleaved by thrombin. MBP tag vectors can be combined with any of the listed below linearized baculovirus DNA vectors, i.e. ProEasy™, ProGreen™, ProFold™-ER1, ProFold™-C2, ProFold™-PDI and ProFold™-0. 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.
 
  Examples
 
  1.Overexpressed cytoplasmic protein forms inclusion bodies. Insert gene encoding your target protein into pAB-MBP™ plasmid transfer vector in-frame with the MBP tag. Co-transfect the resulting construct with ProFold™-C2 linearized baculovirus DNA, select recombinant clones.
  2.Overexpressed secreted or transmembrane protein forms inclusion bodies. Follow our recommendations for Secreted proteins or Transmembrane proteins.
 
  Relevant products
 
Plasmid transfer vectors                             Click on the product name for technical information!
Cat # Product Specification Unit size Price*
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™-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
A7 ProFold™-PDI Overexpressed PDI facilitates folding of cysteine-rich proteins
High level of target protein expression from the same genome as PDI
25 μl, 2.5 μg
5 transfections
$465.00
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 $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
 
  *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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