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Sigma-Aldrich

Calcium Phosphate Transfection Kit

Most cost effective transfection reagent kit for transient and stable transfection of DNA into mammalian cells

Synonym(s):

Gene delivery

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About This Item

EC Number:
233-140-8
UNSPSC Code:
12352200
NACRES:
NA.85

grade

for molecular biology

Quality Level

200

form

solution

usage

 kit sufficient for 160 transfections (6 cm dishes)
 kit sufficient for 400 transfections (3.5 cm dishes)
 kit sufficient for 80 transfections (10 cm dishes)

technique(s)

transfection: suitable

shipped in

dry ice

storage temp.

−20°C

General description

Calcium phosphate transfection is a commonly used method for the introduction of DNA into eukaryotic cells. This technique has been used to obtain both transient and stable transfections in a wide variety of cell types.

Application

Calcium Phosphate Transfection Kit has been used:
  • to enable transfection
  • to transfect Hek293T cells
  • to transfect H29D cells

Suitable for transient and stable transfection of DNA into cultured mammalian cells. The following cells have successfully been transfected using the calcium phosphate method:

BAEC
Bowels melanoma cells
CHO K1
COS-7
Fibroblasts (human embryonic, neo derm)
HEK293
Huh 7
IMR-90
LLC (Lewis Lung Carcinoma)
NIH3T3
PC-12
PCI-13
SH-Sy5Y
SK-Hep-1
T47D

Features and Benefits

  • Suitable for transient and stable transfection
  • Reproducible for a wide range of cell types
  • Widely referenced
  • Inexpensive

Components

The Calcium Phosphate Transfection Kit contains:
5 ml 2.5M CaCl2 (C2052)
25 ml 2x HEPES Buffered Saline (H1012)
25 ml molecular biology grade water (W4502)

Principle

The procedure is based on slow mixing of HEPES-buffered saline containing sodium phosphate with a CaCl2 solution containing the DNA. A DNA-calcium phosphate co-precipitate forms, which adheres to the cell surface and is taken up by the cell, presumably by endocytosis.

Other Notes

The protocol was developed for transient transfection of CHO cells using pSV40-CAT plasmid as a reporter gene.

Pictograms

Exclamation mark
GHS07

Signal Word

Warning

Hazard Statements

H319

Hazard Classifications

Eye Irrit. 2

Storage Class Code

12 - Non Combustible Liquids

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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A new technique for the assay of infectivity of human adenovirus 5 DNA.
F L Graham et al.
Virology, 52(2), 456-467 (1973-04-01)
A Calcium Phosphate Transfection
Ausubel, F. et al (ed.)
Short Protocols in Molecular Biology, 3rd, 9-9 (1995)
Munjin Kwon et al.
Methods in molecular biology (Clifton, N.J.), 1018, 107-110 (2013-05-18)
The calcium phosphate transfection is a widely used method for introducing foreign DNA plasmids into cells. Mechanisms underlying this transfection method are not yet defined; however, DNA-calcium phosphate precipitates are internalized by the cells and DNA is efficiently expressed in
M Wigler et al.
Cell, 14(3), 725-731 (1978-07-01)
Previous studies from our laboratories have demonstrated the feasibility of transferring the thymidine kinase (tk) gene from restriction endonuclease-generated fragments of herpes simplex virus (HSV) DNA to cultured mammalian cells. In this study, high molecular weight DNA from cells containing
Knock-down of SOX11 induces autotaxin-dependent increase in proliferation in vitro and more aggressive tumors in vivo
Conrotto P, et al.
Molecular Oncology, 5(6), 527-537 (2011)
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Articles

Introduction to Cell Transfection

Transfection is the introduction of DNA, RNA, or proteins into eukaryotic cells and is used in research to study and modulate gene expression. Thus, transfection techniques and protocols serve as an analytical tool that facilitates the characterization of genetic functions, protein synthesis, cell growth and development.

How Transfection Works

This brief webinar provides an overview of what transfection is and the methods that are used to introduce DNA or RNA into eukaryotic cells.

Protocols

Calcium Phosphate Transfection Kit Protocol

Calcium phosphate transfection is a common method for the introduction of DNA into eukaryotic cells. This protocol can be optimized for use with a wide variety of cell types.

Our team of scientists has experience in all areas of research including Life Science, Material Science, Chemical Synthesis, Chromatography, Analytical and many others.

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