Ordering EF5 and α-EF5 Antibodies
Please following the steps below to place your order for EF5.
- Read the information printed below to calculate required quantities. More detailed information is located in the references section . For any remaining questions please contact Cameron Koch (firstname.lastname@example.org).
EF5 Amount Calculation Guide
Please see the review with specific information about how EF5 and its detecting antibodies are used.
EF5 (MW of drug is 302) is made at no charge by the NCI (please acknowledge this in any publications) but it is very expensive to make - probably $5 per mg or so. Thus, we ask investigators to calculate the minimum amount needed (e.g. ~ 1 or 2 (for double injection) mg for a mouse and 10 mg for a rat; plus at least 50 mgs extra to make competed stain and rinse solutions). EF5 is only supplied for experimental use - chemical, tissue culture, or animal. NOT FOR HUMAN USE.
You need about 50-100 µl of anti-EF5 antibody (ELK3-51) at 75 µg/ml for a typical section or flow sample. The Service Center runs under non-profit financial conditions and the antibodies (which are already conjugated with biotin or fluorochromes, eliminating secondary staining) cost $1.30 per microgram.
Typically we would send out a kit with perhaps 50-100 mg of EF5, 3mls of Regular Stain and 1ml of Competed Stain antibodies, premixed in an albumin carrier solution at the ready-to-use concentration of 75 µg/ml. We have these in Alexa488, Cy3, Cy5, and Biotin. Larger quantities of antibody are shipped at 1 mg/ml in PBS with 25% glycerol (to facilitate freezing and efficient recovery after re-thawing).
There are quite a few references and bits of information elsewhere on this website.
We usually make a tail-vein injection using 10 mM drug in 5% dextrose (=glucose) + 2.4% ethanol, with the amount (in mls) equal to 1/100 of the animals weight, in grams (0.25 mls for a 25 g mouse). So if the drug were uniformly distributed and the mice were made of water, the initial drug concentration would be 10mM/100 or 100 µM. We use the same concentration for in vitro studies (100 µM, typically for 3 hrs).
To make the EF5 injection solution: make up some 5% glucose (use tissue culture or USP grade). You can then dissolve all the EF5 in only the ethanol component with modest swirling (perhaps assisted by heat from running hot water) and end by adding the glucose solution to it.
For example, to make 10 ml, you need 30 mg of EF5 - add to a 15 ml polypropylene tube (don't use polystyrene). 2.4% of 10 is 0.24 so add 0.24 g (by weight) of Pharmco USP 100% ethanol. Cap the tube and gently swirl in hot water to completely dissolve the EF5. Then add the glucose solution quite quickly to the 10 ml mark. Filter sterilize - keep at room temperature covered in foil to protect from light.
We can also supply small quantities of EF3 and its associated antibodies (ELK5-A8). EF3 costs $10 per milligram, and the anti-EF3 antibodies are only available with Cy3 and Cy5 fluorochromes.