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A) Pull-down of mCherry from a mixture of GFP, mCherry and mTagBFP. B) IP of mCherry 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. Zoom

A) Pull-down of mCherry from a mixture of GFP, mCherry and mTagBFP. B) IP of mCherry 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.

protag-HiPur RFP Agarose Beads, sample

Camelid single-domain antibody (sdAb) covalently linked to 4% agarose beads.
Cat. No.: 89200S
Quantity:  200 µl

Delivery Time: usually 1-7 working days

€148.00
Excl. 19% Tax, excl. Shipping Cost

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Product description

Host llama/ alpaca
Antibody Type recombinant, produced in E.coli
Isotype single-domain antibody
Clone 2B12
Immunogen RFP
Purification affinity chromatography
Formulation 50 % slurry in PBS containing 20 % Ethanol
Storage store at 2 – 8 °C
Tested species reactivity dsRed1, dsRed2, mCherry, mOrange2, mRFP, mScarlet-i, tdTomato (does not cross-react with Dendra2, Dronpa, tdEOS, mEOS3.2, mRuby3, mTFP, GFP, mTagBFP or their most common derivatives)
Intended use Research use only
Support 4 % cross-linked agarose, bead size 50-150 μm
Coating sdAb anti-RFP clone 2B12
Capacity > 4 μg RFP per μl of packed beads
Buffer Compatibility • Common buffer substances at pH 5 to 9
• 2% Triton X-100, 1% Tween-20, 1% NP-40, 1% CHAPS, 1% Deoxycholate, 0.1% SDS
• 4 M NaCl, 2 M KCl, 1 M MgCl2, 100 mM EDTA
• 4 M urea
• 10 mM DTT, 10 mM 2-Mercaptoethanol
• RNAse A, DNAse I, Benzonase, protease inhibitors

Applications

Tested applications Tested dilutions
Immunoprecipitation (IP) yes

Background

Details

protag-HiPur RFP 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 RFP Agarose Beads thus feature high affinity and superior capacity for RFP fusion proteins while showing negligible non-specific background. protag-HiPur GFP Agarose Beads immobilize a wide range of RFP derivatives, such as most common red fluorescent proteins like mRFP, mCherry, dsRed, tdTomato and mScarlet. It does not cross-react with GFP or mTagBFP/mTagRFP derivatives. protag-HiPur RFP Agarose Beads are compatible not only with physiological buffers but also with high stringency buffers. With protag-HiPur RFP Agarose Beads the binding and washing conditions can be adjusted to the experimental needs.

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