protag-HiPur GFP Agarose Beads
A) Pull-down of GFP from a mixture of GFP, mCherry and mTagBFP. B) IP of GFP from HeLa lysate. In/Ft: 1/1000 of input and non-bound material. E: Eluate from 1 µl of beads. C) Control experiment using functionalized beads lacking sdAbs.
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A) Pull-down of GFP from a mixture of GFP, mCherry and mTagBFP. B) IP of GFP from HeLa lysate. In/Ft: 1/1000 of input and non-bound material. E: Eluate from 1 µl of beads. C) Control experiment using functionalized beads lacking sdAbs.
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Key Features
Camelid single-domain antibody (sdAb) covalently linked to 4% agarose beads.
Product description
| Quantity | 2 ml |
|---|---|
| Antibody Type | Recombinant, produced in E.coli |
| Host | Llama/alpaca |
| Isotype | Single-domain antibody |
| Application | IP |
| Purification | Affinity chromatography |
| Reactivity | Cerulean, Citrine, EGFP, Emerald GFP, Sirius, Superecliptic pHluorin, Superfold GFP, acGFP, eCFP, eYFP, mEGFP (A206K), mEGFP (L221K), mTurquoise 1/2, mVenus, paGFP, tSapphire |
| No reactivity | Dendra2, Dronpa, tdEOS, mEOS3.2, mRuby3, mTFP, mCherry, mRFP, dsRed, mTagBFP or their most common derivatives |
| Storage | 2-8°C |
| Intended use | Research use only |
| Clone | 1H1 |
| Immunogen | GFP |
| Formulation | 50% slurry in PBS containing 20% Ethanol |
| Capacity | > 4 µg GFP per µl of packed beads |
| Coating | sdAb anti-GFP clone 1H1 |
| Support | 4% cross-linked agarose, bead size 50-150 µm |
| Buffer compatibility |
|
Applications
| Tested applications | Tested dilutions |
|---|---|
| Immunoprecipitation (IP) | yes |
Background
protag-HiPur GFP Agarose Beads are based on a high-affinity single-domain antibodies (sdAb) that are covalently immobilized on 4% cross-linked agarose beads. The sdAbs are attached via a flexible linker which guarantees a high accessibility of the sdAbs and largely eliminates batch-to-batch variations. Due to the single-chain nature of sdAbs and their covalent attachment, no "leakage" of light and heavy chains from IgGs is observed during elution with SDS sample buffer. protag-HiPur GFP Agarose Beads thus feature high affinity and superior capacity for GFP fusion proteins while showing negligible non-specific background. protag-HiPur GFP Agarose Beads immobilize a wide range of GFP derivatives, such as GFP, mEGFP, superfolder GFP, Emerald GFP and most common CFP and YFP variants. It does not cross-react with mCherry, mRFP, dsRed, mTagBFP or their most common derivatives. protag-HiPur GFP Agarose Beads are compatible not only with physiological buffers but also with high stringency buffers. With protag-HiPur GFP Agarose Beads the binding and washing conditions can be adjusted to the experimental needs.
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