Agilent Technologies Vitality hrGFP Subcellular Localization Vectors User Guide
Summary
These subcellular localization vectors are essential research tools designed to track the movement and precise location of proteins within key cell organelles, including the mitochondria, nucleus, Golgi apparatus, and peroxisomes. This comprehensive manual provides expert guidance on the entire process, covering material handling, electroporation/transfection protocols, and utilizing Cre-mediated recombination techniques. It is intended for molecular biologists and researchers conducting advanced cell biology studies to accurately visualize organelle dynamics in living cells.
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Vitality hr GFP Subcellular Localization Vectors phr GFP-Nuc, phr GFP-Mito, phr GFP-Golgi, and phr GFP-Perox
Instruction Manual Catalog #240042 (phr GFP-Mito) #240043 (phr GFP-Nuc) #240044 (phr GFP-Golgi) #240063 (phr GFP-Perox) Revision D
Research Use Only. Not for Use in Diagnostic Procedures. 240042-12
Page Summary Contents For Agilent Technologies Vitality hrGFP Subcellular Localization Vectors User Guide
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Vitality hr GFP Subcellular Localization Vectors phr GFP-Nuc, phr GFP-Mito, phr GFP-Golgi, and phr GFP-Perox Instruction Manual Catalog #240042 (phr GFP-Mito) #240043 (phr GFP-Nuc) #240044 (phr GFP-Go...
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LIMITED PRODUCT WARRANTY This warranty limits our liability to replacement of this product. No other warranties of any kind, express or implied, including without limitation, implied warranties of mer...
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VITALITY HRGFP SUBCELLULAR LOCALIZATION VECTORS phr GFP-Nuc, phr GFP-Mito, phr GFP-Golgi, and phr GFP-Perox CONTENTS Materials Provided ...................................................................
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Vitality hr GFP Subcellular Localization Vectors phr GFP-Nuc, phr GFP-Mito, phr GFP-Golgi, and phr GFP-Perox MATERIALS PROVIDED Material provided Concentration Quantity phr GFP-Mito vector (Catalog #2...
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NOTICES TO PURCHASER CMV PROMOTER The use of the CMV Promoter is covered under U.S. Patent Nos. 5,168,062 and 5,385,839 owned by the University of Iowa Research Foundation and licensed FOR RESEARCH US...
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from defects of the Products, including, but not limited to, (a) any implied warranty or merchantability or fitness for a particular purpose, (b) any implied warranty arising from course of performanc...
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INTRODUCTION The green fluorescent protein (GFP) has become an extremely versatile tool for tracking and quantifying biological entities in the fields of biochemistry, molecular and cell biology, as w...
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Description of the Vectors The hr GFP-based subcellular localization vectors are derived from the vector p Exchange-1, and thus take advantage of Cre-mediated site-specific recombination to allow quic...
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The phr GFP-Mito Vector P CMV p UC ori mito signal phr GFP-Mito hr GFP 4.3 kb ampicillin SV40 p A Lox P f1 ori Feature Nucleotide Position CMV promoter 1–597 mitochondrial localization signal 606–713 ...
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The phr GFP-Nuc Vector P CMV p UC ori hr GFP phr GFP-Nuc 4.3 kb ampicillin SV40 NLS SV40 p A Lox P f1 ori Feature Nucleotide Position CMV promoter 1–597 hr GFP ORF 606–1322 SV40 nuclear localization s...
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The phr GFP-Golgi Vector P CMV p UC ori phr GFP-Golgi hr GFP 4.3 kb ampicillin SV40 p A Lox P f1 ori Feature Nucleotide Position CMV promoter 1–597 sialyltransferase transmembrane domain (TMD) 606–710...
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The phr GFP-Perox Vector P CMV p UC ori hr GFP phr GFP-Perox 4.2 kb ampicillin perox signal SV40 p A Lox P f1 ori Feature Nucleotide Position CMV promoter 1–597 hr GFP ORF 606–1322 peroxosomal localiz...
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OVERVIEW OF CRE-MEDIATED SITE-SPECIFIC RECOMBINATION The Agilent Vitality hr GFP subcellular localization vectors utilize Cre- mediated site-specific recombination to allow the quick and efficient dir...
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MODULE DESCRIPTIONS The Vitality hr GFP vectors allow for the introduction of a desired eukaryotic resistance gene into the core expression vectors. Each module, shown in Figure 6, contains a Lox P si...
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CRE-MEDIATED RECOMBINATION PROTOCOL 1. To prepare the recombination reaction, add the following components to a 0.5-ml microcentrifuge tube: X μl of Vitality hr GFP vector containing the gene of inter...
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MAMMALIAN CELL TRANSFECTION Prepare enough DNA of appropriate purity for the mammalian cell transfection procedure to be carried out. Protocols for transfection of mammalian cell lines can be found in...
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TROUBLESHOOTING Observation Suggestion Very few colonies following transformation with Transformation efficiency is inhibited by Cre recombinase that is not Cre-recombined vector denatured by heat tre...
Manual Details
| Brand | Agilent Technologies |
|---|---|
| Pages | 17 |
| File Size | 323.59 KB |
| Published | May 13, 2026 |
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Frequently Asked Questions
What should I do if I see very few colonies after transformation?
Heat the recombination reaction at 65°C for 20 minutes to denature the Cre recombinase.
Which vector is used for Golgi localization?
The phrGFP-Golgi vector uses catalog number #240044.
Can these vectors be used in diagnostic procedures?
No, the product is strictly designated for 'Research Use Only' and not for diagnostic procedures.
How do I prepare LB Agar media?
Add 10g NaCl, 10g tryptone, 5g yeast extract, and 20g agar to water, adjust pH to 7.0, and autoclave.