Sapphire Insect Transfection Kit

SapphireTM Insect Transfec-
tion Kit
I nsect cells and lytic baculoviruses

provide a proven method for high-
level expression of full-length mam-
5. Mix well and add the mixture from
Step 4 dropwise to cells from Step 2.
For negative control, use Sapphire™ F or Research Use Only. Not for
Diagnostic or Therapeutic Use.
malian proteins. Autographa californi- DNA alone without any vector DNA. Purchase does not include or carry
any right to resell or transfer this
ca nuclear polyhedrosis virus (AcNPV) For positive control, use pVL1392- product either as a stand-alone
is used to infect cultured insect cells XylE (Cat. No. ABP-BVP-10001) in product or as a component of another
(e.g. Spodoptera frugiperda). Expres- combination with Sapphire™ DNA. product. Any use of this product other
sion of the highly abundant polyhedrin 6. Incubate the plates at 27°C for than the permitted use without the
gene is nonessential in tissue culture 4 hours. After that time, remove medi- express written authorization of Allele
and its strong promoter can be used um and replace it with fresh TNM-FH Biotech is strictly prohibited
for the transcription of foreign genes. medium containing 10% fetal bovine
The polyhedrin promoter is maximally serum (Cat. No. ABP-MED-10001).
expressed very late stage in infection 7. After 5 days, collect the superna- Website: www.allelebiotech.com
when the lytic virus kills the host cells, tant of all three plates and infect fresh Call: 1-800-991-RNAi/858-587-6645
resulting in high levels of expression cells to amplify the virus. Optionally, (Pacific Time: 9:00AM~5:00PM)
even for certain toxic proteins. In ad- lyse the transfected cells and check Email: [email protected]
dition, many post-translational modifi- for expression of your protein of
cations similar to those in mammalian interest.
cells are made in insect cells and pro-
teins unable to be expressed in E. coli
have been successfully expressed Box 1 | Product Summary
in the insect cell system. The Sap-
Catalogue Number ABP-BVD-10003
phire™ Insect Transfection Kit is ideal
for easy and efficient transfection of Component 1 bottle of Transfection Buffer A, 25ml
DNAs into insect cells. It is optimized 1 bottle of Transfection Buffer B, 25ml
for use with Sapphire™ Baculovirus Storage Store at +4°C.
DNA and various transfer vectors.
Stability The transfection buffers are stable for six months
when stored properly.
Protocol
Transfection Using the Sap-

phire™ Transfection Kit
1. Seed 2 x 106 Sf9 (Cat. No. ABP- Box 2 | Related Products
CEL-10001) or Sf21 (Cat. No. ABP-
CEL-10002) cells onto each 60mm Product Catalogue No.
tissue culture plate. Allow cells to
Sapphiretm Baculovirus DNA (5µg) ABP-BVD-10001
attach firmly which takes usually 5 to
10 min. Sapphire™ Baculovirus DNA and Transfection ABP-BVD-10002
Kit
Note: Prepare three plates for each
co-transfection: one for the positive Sf9 Cells (in culture, 107 cells) ABP-CEL-10002
control, one for the negative control, Sf9 Cells (frozen, 10 cells)
7
ABP-CEL-10006
and one for the recombinant plasmid
Sf21 Cells (in culture, 107 cells) ABP-CEL-10003
of interest.
2. Remove medium and replace it Sf21 Cells (frozen, 10 cells)
7
ABP-CEL-10007
with 1 ml of sterile Transfection Buffer T.ni Cells (in culture, 10 cells)
7
ABP-CEL-10005
A. T.ni Cells (frozen, 107 cells) ABP-CEL-10008
3. Mix 0.5 μg of Sapphire™ or other
TNM-FH Insect Culture Medium ABP-MED-10001
baculoviral DNA and 2μg of recom-
binant baculovirus transfer vector Serum Free Insect Culture Medium ABP-MED-10002
containing gene of interest in a sterile Serum Free, Met Free Insect Culture Medium ABP-MED-10003
microfuge tube.
Grace’s Insect Culture Medium ABP-MED-10004
4. Let mixture from Step 3 sit for 1
min at room temperature before add- pVL-1392-XyIE Control Vector ABP-BVP-10001
ing 1 ml of sterile Transfection Buffer Insect Lysis Buffer 5X (10ml) ABP-BUF-10010
B.
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Sapphire Insect Transfection Kit

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Available Formats

SapphireTM Insect Transfec-

I nsect cells and lytic baculoviruses

Transfection Using the Sap-

Allele Biotech-Introducing Cost Effectiveness to Research

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