Thermo Fisher Scientific CellSensor Lentiviral Vectors User guide

Brand: Thermo Fisher Scientific

Model: CellSensor Lentiviral Vectors

Type: User guide

CellSensor® Lentiviral Vectors User Guide

Cat. nos. K1109, K1110, K1115, K1116, K1117, K1118, K1119, K1121, K1124, K1125, K1126,

K1128, K1135, K1132, K1133, K1164, K1193, K1194, K1195, K1196, and K1242

Lit. No. O13404.pps Rev Date: 31 May 2006 Shipping Condition: Dry Ice Storage: –20°C

For Technical Support for this or any other Invitrogen Drug Discovery Solutions Products, dial 760 603 7200, extension 40266

Invitrogen Corporation • 1600 Faraday Avenue • Carlsbad, CA 92008 • Phone: 760 603 7200 • FAX: 760 602 6500 • www.invitrogen.com

TABLE OF CONTENTS

1.0 MATERIALS SUPPLIED ............................................................................................................................................................ 1

2.0 MATERIALS REQUIRED, BUT NOT SUPPLIED....................................................................................................................... 1

3.0 VIRAPOWER™ LENTIVIRAL EXPRESSION SYSTEM OVERVIEW ......................................................................................... 2

3.1 Description ................................................................................................................................................................. 2

3.2 How the ViraPower™ Lentiviral Expression System Works ................................................................................... 2

4.0 LIPOFECTAMINE™ 2000 TRANSFECTION REAGENT ............................................................................................................. 3

4.1 Description .................................................................................................................................................................. 3

4.2 How Lipofectamine™ 2000 Transfection Reagent Works ....................................................................................... 3

5.0 ENHANCER ELEMENTS AVAILABLE ..................................................................................................................................... 4

6.0 BIOSAFETY LEVEL AND HANDLING ..................................................................................................................................... 5

7.0 PURCHASER NOTIFICATION .................................................................................................................................................. 6

1.0 MATERIALS SUPPLIED

Description: CellSensor® Lentiviral Vectors

Shipping Condition: Dry Ice

Storage Condition: –20°C

Quantity: 10 µg

2.0 MATERIALS REQUIRED, BUT NOT SUPPLIED

Reagents Recommended Source Cat. no.

ViraPower™ Bsd Lentiviral Support Kit Invitrogen K4970-00

Lipofectamine™ 2000 Transfection Reagent Invitrogen 11668-027

11668-019

Invitrogen •CellSensor® Lentiviral Vectors User Guide • O13404pps • 09/05 Page 2 of 6

For Technical Support for this or other Invitrogen Drug Discovery Solutions Products, dial 760 603 7200, extension 40266

Invitrogen Corporation • 1600 Faraday Avenue • Carlsbad, CA 92008 • Phone: 760 603 7200 • FAX: 760 602 6500 • www.invitrogen.com

3.0 VIRAPOWER™ LENTIVIRAL EXPRESSION SYSTEM OVERVIEW

3.1 Description

CellSensor® Lentiviral Vectors may be used in combination with the ViraPower™ Lentiviral Expression System for

viral transduction of mammalian cells or analogous to a standard vector for transfection of mammalian cells with

traditional lipid-based transfection techniques.

Many mammalian cell types and experimental situations create challenges for standard transfection experiments.

The ViraPower™ Lentiviral Expression System overcomes these challenges with efficient viral delivery to both

dividing and non-dividing cells. Lentivirus is ideal for analysis of long-term gene expression, even in terminally

differentiated or growth-arrested cells. Use this system to analyze gene expression beyond the limitations of

traditional transfection reagents or other viral systems. The ViraPower™ Lentiviral Expression System provides

the high levels of stable gene expression necessary for valid results in virtually any cell line.

3.2 How the ViraPower™ Lentiviral Expression System Works

High-level stable gene expression is easily achieved with the ViraPower™ Lentiviral Expression system (Figure 1).

Co-transfect the vector containing your gene of interest along with the optimized ViraPower™ Lentiviral

Packaging Mix into 293FT producer cells. The ViraPower™ Packaging Mix provides a pre-optimized mix of

plasmids that produce the lentiviral packaging proteins (gag/pol and rev) and the VSV-G envelope protein. The

VSV-G envelope gives the particles produced the ability to transduce a broad spectrum of mammalian cells. In 24-

48 hours harvest the supernatant, which now has replication-incompetent viral particles that contain your gene of

interest. Since the packaging proteins are provided in trans, these plasmids are not part of the final viral particles.

You can use this supernatant to efficiently transduce any mammalian cell. Each transfection produces sufficient

supernatant for multiple transduction experiments.

Detailed protocols, citations, and additional product information for the ViraPower™ Lentiviral Expression

System can be found at www.invitrogen.com.

Invitrogen •CellSensor® Lentiviral Vectors User Guide • O13404pps • 09/05 Page 3 of 6

For Technical Support for this or other Invitrogen Drug Discovery Solutions Products, dial 760 603 7200, extension 40266

Invitrogen Corporation • 1600 Faraday Avenue • Carlsbad, CA 92008 • Phone: 760 603 7200 • FAX: 760 602 6500 • www.invitrogen.com

4.0 LIPOFECTAMINE™ 2000 TRANSFECTION REAGENT OVERVIEW

4.1 Description

Lipofectamine™ 2000 Reagent is a cationic lipid that offers the highest transfection efficiencies of the

widest variety of cell lines and high protein expression levels or gene inhibition (RNAi). It is the leading

transfection reagent for simple, effective, and optimal nucleic acid delivery. Lipofectamine™ 2000 Reagent

provides all of the following:

• Exceptional transfection efficiency in a broad range of cell lines and high levels of recombinant

protein expression

• Superior performance for siRNA delivery

• Proven efficacy in the presence of serum—eliminates the need to change media following transfection

• A reagent proven to work in high-throughput applications

• A reliable reagent for establishing stable cell lines

4.2 How Lipofectamine™ 2000 Transfection Reagent Works

Lipofectamine™ 2000 Reagent offers high transfection efficiencies for a broad range of cell types,

including adherent and suspension cell lines. This means you can choose the most appropriate cell line

for gene function studies or target validation. For cell lines commonly used for protein expression and

cell-based functional assays, including COS-7, CHO and HEK 293T, efficiencies are often greater than

90%. Lipofectamine™ 2000 Reagent works effectively with many primary cell lines and other difficult to

transfect cell lines. In addition to its versatility with diverse cell lines, Lipofectamine™ 2000 provides high

transfection efficiencies using media with or without serum. For cells sensitive to the absence of serum,

add DNA:lipid complexes directly to cells in serum-containing medium.

For gene silencing, Lipofectamine™ 2000 Reagent’s high efficiency transfections leads to the high levels of

gene knockdown needed to achieve successful results. Simply mix Lipofectamine™ 2000 Reagent with

nucleic acid, add to cell culture, and express protein or knockdown genes. The simplicity and speed

combined with high transfection efficiency make Lipofectamine™ 2000 Reagent ideal for transient protein

expression or high throughput RNAi. Transfection conditions can be easily established for automated or

robotic systems used in such applications.

Protocols, citations, and additional product information for Lipofectamine™ 2000 can be found in the Cell

Lines Database at www.invitrogen.com.

Invitrogen •CellSensor® Lentiviral Vectors User Guide • O13404pps • 09/05 Page 4 of 6

For Technical Support for this or other Invitrogen Drug Discovery Solutions Products, dial 760 603 7200, extension 40266

Invitrogen Corporation • 1600 Faraday Avenue • Carlsbad, CA 92008 • Phone: 760 603 7200 • FAX: 760 602 6500 • www.invitrogen.com

5.0 ENHANCER ELEMENTS AVAILABLE

Cat. no. Product Element Transcription

Facto

Pathway

K1109 pLenti-bsd/NFAT-bla Vector Nuclear Factor of Activated

T-Cells (NFAT) Response NFAT PKC and Ca2+/Calcineurin

K1115 pLenti-bsd/ISRE-bla Vector Interferon Stimulated

Response Element (ISRE) STAT1/STAT2 Proliferation/Inflammation

Jak/STAT

K1116 pLenti-bsd/SIE-bla Vector SIE STAT3 Proliferation/Inflammation

Jak/STAT

K1117 pLenti-bsd/SBE-bla Vector Smad Binding Element (SBE) Smads Transforming Growth

Factor Beta (TGF-β)

K1118 pLenti-bsd/AP-1-bla Vector Activator Protein 1 (AP-1)

Response Element c-jun/c-fos JNK

K1119 pLenti-bsd/c-fos-bla Vector C-fos Promoter (SIE SRE CRE) STAT3,

ATF2/CREB,

JNK/p38/MAPK and

Jak/STAT

K1110 pLenti-bsd/CRE-bla Vector cAMP Response Element (CRE) ATF2/CREB JNK/P38 and PKA

K1121 pLenti-bsd/SRE-bla Vector Serum Response Element (SRE) Elk-1/SRF MAPK/JNK

K1124 pLenti-bsd/HRE-bla Vector Hypoxia Responsive

Elements (HRE) HIF Angiogenesis

K1125 pLenti-bsd/myc-bla Vector E Box DNA Binding Element Myc/Max Cell Proliferation

K1126 pLenti-bsd/LEF/TCF-bla Vector Lymphoid Enhancing Factor

(LEF)/T Cell Factor (TCF) DNA TCF/LEF Wnt-Frizzled

K1128 pLenti-bsd/NFκB-bla Vector Nuclear Factor of B (NFκB)

Cells Res

onse Element NFκB NFκB

K1135 pLenti-bsd/Gli-bla Vector Gli Hedgehog Hedgehog

K1164 pLenti-bsd/GAS-bla Vector Gamma-Interferon

Activation Sequence (GAS) STAT1 Proliferation/Inflammation

Jak/STAT

K1193 pLenti-bsd/p53RE-bla Vector p53 Response Element p53 Cell Growth/Apoptosis

K1194 pLenti-bsd/FHRE-bla Vector Forkhead Response Element FOXO3 PI3K/Akt/Forkhead

K1195 pLenti-bsd/C.EBP-bla Vector C/EBP Response Element C/EBPα C/EBP (differentiation)

K1196 pLenti-bsd/dhfr(E2F) Vector E2F/DP1 Response Element E2F/DP1 RB-E2F/cell cycle

K1242 pLenti-bsd/STAT6-bla Vector STAT6 STAT6 JAK/STAT (IL-4)

6.0 BIOSAFETY LEVEL AND HANDLING

The pLenti vectors should be handled at Biosafety Level 1 and virus produced by this method should be handled

at Biosafety Level 2. If the vector is used analogous to a standard vector, all material may be used at Biosafety

Level 1.

Invitrogen •CellSensor® Lentiviral Vectors User Guide • O13404pps • 09/05 Page 5 of 6

For Technical Support for this or other Invitrogen Drug Discovery Solutions Products, dial 760 603 7200, extension 40266

Invitrogen Corporation • 1600 Faraday Avenue • Carlsbad, CA 92008 • Phone: 760 603 7200 • FAX: 760 602 6500 • www.invitrogen.com

7.0 PURCHASER NOTIFICATION

Limited Use Label License No. 5: Invitrogen Technology

The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and

components of the product in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The

buyer cannot sell or otherwise transfer (a) this product (b) its components or (c) materials made using this product or its

components to a third party or otherwise use this product or its components or materials made using this product or its

components for Commercial Purposes. The buyer may transfer information or materials made through the use of this product

to a scientific collaborator, provided that such transfer is not for any Commercial Purpose, and that such collaborator agrees in

writing (a) not to transfer such materials to any third party, and (b) to use such transferred materials and/or information

solely for research and not for Commercial Purposes. Commercial Purposes means any activity by a party for consideration

and may include, but is not limited to: (1) use of the product or its components in manufacturing; (2) use of the product or its

components to provide a service, information, or data; (3) use of the product or its components for therapeutic, diagnostic or

prophylactic purposes; or (4) resale of the product or its components, whether or not such product or its components are

resold for use in research. Invitrogen Corporation will not assert a claim against the buyer of infringement of patents owned or

controlled by Invitrogen Corporation which cover this product based upon the manufacture, use or sale of a therapeutic,

clinical diagnostic, vaccine or prophylactic product developed in research by the buyer in which this product or its

components was employed, provided that neither this product nor any of its components was used in the manufacture of such

product. If the purchaser is not willing to accept the limitations of this limited use statement, Invitrogen is willing to accept

return of the product with a full refund. For information on purchasing a license to this product for purposes other than

research, contact Licensing Department, Invitrogen Corporation, 1600 Faraday Avenue, Carlsbad, California 92008. Phone

(760) 603-7200. Fax (760) 602-6500. Email: o[email protected]

Limited Use Label License No: 51 Blasticidin and the Blasticidin Selection Marker

Blasticidin and the blasticidin resistance gene (bsd) are the subject of U.S. Patent No. 5,527,701sold under patent license for

research purposes only. Inquiries for commercial use should be directed to: Kaken Pharmaceutical Company, Ltd., Bunkyo

Green Court, Center Office Building, 19-20 Fl, 28-8 Honkomagome 2-chome, Bunkyo-ku, Tokyo 113-8650, Japan, Tel: 81 3-

5977-5008; Fax: 81 3-5977-5008

Limited Use Label License No: 108 Lentiviral Technology

The Lentiviral Technology (based upon the lentikat™ system) is licensed from Cell Genesys, Inc., under U.S. Patent Nos.

5,686,279; 5,834,256; 5,858,740; 5,994,136; 6,013,516; 6,051,427; 6,165,782 and 6,218,187 and corresponding patents and

applications in other countries for internal research purposes only. Use of this technology for gene therapy applications or

bioprocessing other than for non-human research use requires a license from Cell Genesys (Cell Genesys, Inc. 342 Lakeside

Drive, Foster City, California 94404). The purchase of this product conveys to the buyer the non-transferable right to use the

purchased amount of the product and components of the product in research conducted by the buyer, including non-gene

therapy research and target validation applications in laboratory animals (whether the buyer is an academic or for-profit

entity). The buyer cannot sell or otherwise transfer (a) this product (b) its components or (c) materials made using this product

or its components to a third party or otherwise use this product or its components or materials made using this product or its

components for Commercial Purposes. The buyer may transfer information or materials made through the use of this product

to a scientific collaborator, provided that such transfer is not for any Commercial Purpose, and that such collaborator agrees in

writing (a) not to transfer such materials to any third party, and (b) to use such transferred materials and/or information

solely for research and not for Commercial Purposes. Commercial Purposes means any activity by a party for consideration

and may include, but is not limited to: (1) use of the product or its components in manufacturing; (2) use of the product or its

components to provide a service, information, or data; (3) use of the product or its components for therapeutic, diagnostic or

prophylactic purposes; or (4) resale of the product or its components, whether or not such product or its components are

resold for use in research. Invitrogen Corporation will not assert a claim against the buyer of infringement of the above

patents based upon the manufacture, use or sale of a therapeutic, clinical diagnostic, vaccine or prophylactic product

developed in research by the buyer in which this product or its components was employed, provided that neither this product

nor any of its components was used in the manufacture of such product. If the purchaser is not willing to accept the

limitations of this limited use statement, Invitrogen is willing to accept return of the product with a full refund. For

information on purchasing a license to this product for purposes other than research, contact Licensing Department,

Invitrogen Corporation, 1600 Faraday Avenue, Carlsbad, California 92008. Phone (760) 603-7200. Fax (760) 602-6500.

Invitrogen •CellSensor® Lentiviral Vectors User Guide • O13404pps • 09/05 Page 6 of 6

For Technical Support for this or other Invitrogen Drug Discovery Solutions Products, dial 760 603 7200, extension 40266

Invitrogen Corporation • 1600 Faraday Avenue • Carlsbad, CA 92008 • Phone: 760 603 7200 • FAX: 760 602 6500 • www.invitrogen.com

Limited Use Label License No. 150: GeneBLAzer® Technology

This product and/or its use is the subject of one or more of U.S. Patent Nos. 5,741,657, 5,955,604, 6,291,162, and 6,472,205 and

foreign equivalents licensed to Invitrogen Corporation. The right to use this product for internal research specifically excludes

the right to use this product to identify, discover, and profile compounds that act as a flavor, fragrance or taste-enhancers and

modify a target identified in taste, olfaction, or pheromone detection, which compound does not require FDA approval of an

NDA for claims of safety and efficacy. The right to use methods claimed in the foregoing patents with this product for

research purposes can only be acquired by the use of GeneBLAzer® substrates purchased from Invitrogen Corporation or its

authorized distributors

GeneBLAzer® and CellSensor® is a registered trademark of Invitrogen Corporation.

Lipofectamine™, and ViraPower™ are trademarks of Invitrogen Corporation.

Not intended for any animal or human therapeutic or diagnostic use.

Thermo Fisher Scientific CellSensor Lentiviral Vectors User guide

Brand: Thermo Fisher Scientific

Model: CellSensor Lentiviral Vectors

Type: User guide

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Thermo Fisher Scientific CellSensor Lentiviral Vectors User guide

Thermo Fisher Scientific CellSensor Lentiviral Vectors User guide

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