Formedium products in research into WASP family proteins

Our HL5 Medium and Lo-Flo Medium has been used in the research into WASP family proteins by “The American Society for Cell Biology” and authors Douwe M. Veltman and Robert H. Insall.

The following is a snippet from the report:

Dictyostelium Cell Strains, Constructs, and Culture Conditions

The axenic Dictyostelium discoideum strain AX3 was used as wild type. Cells were cultivated in Petri dishes under HL5 medium (Formedium). The open reading frames of WASP B [DictyBase:DDB_G0272811] and WASP C [DictyBase:DDB_G0283827] were amplified from cDNA using PCR. The full length genes were then cloned into green fluorescent protein (GFP) expression vector pDM448 (Veltman et al., 2009 ). The resulting plasmids were electroporated into Dictyostelium using standard conditions and transformants were selected with 50 μg/ml hygromycin. For double expression with clathrin, the clathrin light-chain gene [DictyBase:DDB_G0277403] was cloned into the monomeric red fluorescent protein (mRFP) shuttle vector pDM413 (Veltman et al., 2009 ). The mRFP-clathrin light-chain fusion cassette was then excised using NgoMIV and cloned into the unique NgoMIV site of the WASP B or WASP C expression vector. To reduce autofluorescence, cells were incubated under Lo-Flo medium (Formedium) for 18 h before imaging.

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Reference: – August 15, 2010. Douwe M. Veltman, Robert H. Insall and Thomas Pollard, Monitoring Editor