What is GFP-nAb?
Optimized by the research team at Allele Biotechnology, GFP-nAb™ is a highly specific GFP (Green Fluorescent Protein) binding protein derived from camelids. It is characterized by a small barrel shaped structure (13 KDa, 2.5nm X 4.5 nm) and a very high stability (stable up to 70°C, functional within 2M NaCl or 0.5% SDS). Experiments have shown that one molecule of GFP-nAb™ binds one molecule of GFP with a dissociation constant (Kd) in the sub nanomolar range. This makes GFP-nAb™ the ideal candidate for a variety of biological assays, some of which are listed below:
- Immunoprecipitation / CO-IP
- Quantitative analysis
- Chromatin Immunoprecipitation (ChIP)
- Superresolution Imaging
GFP-nAb™ binds a large number of commonly used fluorescent proteins derived from the original Aequorea victoria GFP, including EGFP, Venus, Cerulean, and EBFP2 (click here for a full list). Conveniently, it displays no binding affinity to non-jellyfish-derived fluorescent proteins, including the "mFruits" such as mCherry, or Allele's mNeonGreen, mTFP1, mWasabi, and mMaple fluorescent proteins. The GFP-nAb™ Spin Kit offers a simple, consistent protocol to cleanly pull down GFP fusions. Each GFP-nAb™ Spin Kit comes with all required buffers and spin columns to eliminate inconsistent elutions and allow you to perform multiple immunoprecipiation reactions in a matter of minutes.
Complete Pulldown using the GFP-nAb™ Spin Kit.
EGFP-expressing Sf9 (insect) cell lysate contained a total of 16µg of EGFP in total volume of 500µl, determined spectrophotometrically. Following the GFP-nAb™ Spin Kit binding and wash protocols, the protein was eluted in 2 x 50 µl elution buffer (0.2M glycine pH 2.5), pooled, and neutralized with 10µl of 1M Tris base. Equal volumes of lysate input (I), flow-through (FT) after binding to GFP-nAb™ agarose resin, and elution (E) fractions were analyzed by SDS-PAGE followed by Coomassie staining. In this experiment, EGFP was quantitatively removed from the lysate.
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