Why Smart-IP is the smart way to IP

Posted by Bindi M. Doshi, PhD on Mar 26, 2020 1:00:00 PM

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 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

Antibody/Beads (pg/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.


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Topics: Epitope Tag

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