QuantiChrom™ Calcium Assay Kit 500

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Quality level: 200

Qualification: for molecular biology

Sterility: sterile; 0.2 μm filtered

Form: solution

Use

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

Application(s): transfection: adequate

Sent in: dry ice

Storage temperature: −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 transient and stable transfections in a wide variety of cell types.

Request

The calcium phosphate transfection kit has been used:

to allow transfection
to transfect Hek293T cells
to transfect H29D cells

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

  • BASS
  • Intestinal melanoma cells
  • CHO K1
  • COS-7
  • Fibroblasts (human embryonic, neo derm)
  • HEK293
  • Eh 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
  • Cheap

Components

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

Beginning

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

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

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