CAPHOS

Calcium Phosphate Transfection Kit

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

• Synonym(s): Gene delivery
• EC Number: 233-140-8
• UNSPSC Code: 12352200
• NACRES: NA.85

Properties

• grade: Molecular Biology
• Quality Level: 200
• form: solution
• usage: 1 kit sufficient for 160 transfections (6 cm dishes); 1 kit sufficient for 400 transfections (3.5 cm dishes); 1 kit sufficient for 80 transfections (10 cm dishes)
• technique(s): transfection: suitable
• shipped in: dry ice
• storage temp.: −20°C

Description

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.

Safety Information

• Pictograms: GHS07
• Signal Word: Warning
• Hazard Statements: H319
• Precautionary Statements: P264 - P280 - P305 + P351 + P338 - P337 + P313
• Hazard Classifications: Eye Irrit. 2
• Storage Class Code: 12 - Non Combustible Liquids
• Flash Point(F): Not applicable
• Flash Point(C): Not applicable