FAQs
Q. What is Bioform
A.
Q. I’m using my own his-tagged membrane-associated protein(s), should I be concerned about orientation of the protein fragments?
A. BioForm is designed to orient the proteins on the surface as depicted in the scheme shown on the BioForm product page. However, to insure this proper orientation your proteins should be at the membrane proximal end. Proteins tagged at the membrane-distal end may benefit from templating, but will most likely not have the orientation needed for building a pathway.
Q. Can I assemble heterogeneous mixtures of proteins using the template-directed-assembly approach?
A. Yes. Simply mix your histidine-tagged proteins to your desired ratio and add BioForm.
Q. Can I bind accessory factors, adaptor proteins, or scaffold proteins to these BioForm-enabled units?
A. Yes, this will require proteins purified without (or removed) tags. As protocols are developed for the specific pathways, this information will be published on the MIL/PAT websites. We recommend adjusting experimental protocols according to your needs. In most cases, simply adding template at 1/10000th and performing the assay, as stated in the MIL literature, is a good starting composition. However, changing the BioForm reagent:protein ratio and/or the total concentrations of reagents may improve the performance of your proteins in the BioForm-enabled assay.
Q. What is the binding capacity of BioForm?
A. This will depend on the protein size and location and availability of the histidine-tag for binding. One BioForm particle has the capacity to bind many proteins.
Q. What is the concentration of Nickel in BioForm?
A. The template is supplied at 1 nM, and contains 100 µM Ni2+.
Q. How much EDTA can be safely added to the mixture?
A. EDTA and EGTA should not be used in any prepared buffers. Commercial proteins come in buffers with these chemicals but there is no need to perform dialysis. Up to 10 µM has been safely used.
Q. My inhibitors are supplied in DMSO. Can I use DMSO with BioForm?
A. Yes, the template has been employed in the presence of up to DMSO at 5% v/v.
Q. Is the BioForm compatible with detergents?
A. In general, no, but the low concentration of detergents in the buffers of commercially available proteins is compatible with the template. No additional detergent should be added to your assays.
Q. I have access to a Caliper LabChip system, is BioForm compatible with this format?
A. Yes. BioForm has been tested and performed well with this system.