ChromoTek’s unconjugated Nanobodies or VHHs are well characterized and validated for the listed applications. Because the molecular weight of these single-domain antibodies (sdAb) is just about 15 kDa, VHHs are only one tenth the size of a conventional IgG antibody (150 kDa) or less than a third the size of a Fab fragment (50 kDa). In combination with their high chemical and thermal stability plus high affinity, these binding proteins are ideal for multiple types of affinity assays in biochemistry and microscopy, including immunoprecipitation, immunofluorescence, super resolution microscopy, surface plasmon resonance (SPR), biolayer interferometry (BLI), switchSENSE technology, etc.
GFP VHH, biotinylated
p53 C-term VHH
p53 N-term VHH
Ligand capture and Affinity Determination Methods
Antibodies or nanobodies can be used to immobilize proteins on various surfaces, such as microtiter plates, beads or biosensors. Protein-protein interactions and affinity (kon/koff and KD values) can be measured with ligand capture-based assay setups, e.g. by surface plasmon resonance (SPR, BiacoreTM/GE Healthcare), biolayer interferometry (BLI, FortéBio/Pall) or the switchSENSE® technology (Dynamic Biosensors). As a prerequisite for these assays, the affinity of the antibody or Nanobody to the capture ligand has to be high enough to assure ligand binding during the time of the measurement.
ChromoTek’s anti-GFP Nanobody has an extremely high affinity to GFP, with a KD value of approx. 1 pM, as determined by switchSENSE. Note that the other Nanobodies from ChromoTek like the GST, MBP VHHs do have single digit nanomolar KD values, which also allow for high affinity binding in before mentioned applications. Therefore, it is a perfect tool for capturing GFP-tagged proteins for protein-protein interaction studies and affinity determination. Furthermore, its high stability allows regeneration and reuse of the biosensor for multiple measurements.PRODUCTS
Only for research applications, not for diagnostic or therapeutic use!