You have been given the mission to start a new project. A tiny tube is put in your hands and you’re told it contains a potentially interesting protein and your job is to characterize it. After you wipe nervous sweat from your brow, you get down to business. Step one- how to isolate your new buddy? Immunoprecipitation can help with this question! You determine which epitope tag is on your protein and go to mblintl.com to see what you can buy to help this new adventure. You see there are antibodies conjugated to magnetic beads, which help protein pull downs from cell lysates. It's easy, it's fast. Hmmmm- promising.
Smart IP magnetic beads and its corresponding tag antibody are covalently conjugated. The magnetic beads are 1.5 µm in diameter in size. The amount of antibody/bead is listed below:
|Code No.||Immunogen||IgG Volume||Beads volume||Number of Beads||
|D153-11||GFP||160 ug||10 mg||4x10^9||0.04|
|D291-11||His||130 ug||10 mg||4x10^9||0.0325|
|M047-11||Myc||300 ug||15 mg||6x10^9||0.05|
|M132-11||HA||210 ug||10 mg||4x10^9||0.0525|
|M167-11||V5||180 ug||10 mg||4x10^9||0.045|
|M165-11||RFP||140 ug||10 mg||4x10^9||0.035|
|M185-11||DDDDK||360 ug||15 mg||6x10^9||0.06|
|M180-11||HA||250 ug||10 mg||4x10^9||0.0625|
|M075-11||83 ug||10 mg||4x10^9||0.0208|
|M076-11||150 ug||10 mg||4x10^9||0.0375|
|M077-11||150 ug||10 mg||4x10^9||0.0375|
|M081-11||210 ug||10 mg||4x10^9||0.0525|
10 mg beads = 4 x 10^9 beads
In addition, the Smart IP series do not require a pre-clear step. Some labs choose to pre-clear to reduce non-specific binding. To do this, it is best to use an irrelevant antibody. While MBL does not believe a pre-clear step is necessary, if you believe your sample contains some non-specific binding issues, we recommend M081-11 (isotype control beads) for a pre-clear step.
Now you have information. You can plan your experiment, characterize your protein, and make your PI happy.
Watch our super cool Smart-IP video! http://youtu.be/_4kR3Wxlbpk