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FeBABE

Item # Unit Size
F279-10
1 mg

For Research Use Only Products

MSDS
Chemical Name: 1-(p-Bromoacetamidobenzyl) ethylenediamine N,N,N’,N’-tetraacetic acid, acid iron(III)

Appearance:
Yellowish-brown powder
Purity: ≥95.0% (HPLC)
MW:
571.14, C19H21BrFeN3O9

Storage Condition: -20oC
Shipping Condition: ambient temperature


Product Description of BABEs
Bromoacetamidobenzyl-EDTA (BABE) is a chelate labeling reagent that conjugates with sulfhydryl groups. The iron chelate of BABE (FeBABE) is a unique tool for determining the three-dimensional structure of proteins and the binding structures of protein-protein or protein-DNA complexes. BABE adds EDTA moieties to proteins through their sulfhydryl groups. Once attached to a protein, FeBABE cuts a nearby peptide or DNA chain. The cleavage site is within 12 angstroms of the FeBABE binding site. Iron (II)-chelate cleaves a peptide or DNA chain in the presence of hydrogen peroxide. The cleavage reaction completes quickly: 10 seconds to 20 minutes of incubation is sufficient. The size of the cleaved fragment is analyzed with gel electrophoresis such as SDS-PAGE.

Structural Formula:



Labeling Procedure
1. Dialyze the protein solution in conjugation buffer (10-20 mM MOPS, 0.2 M NaCl, 2 mM EDTA, 5% glycerol, pH 8.0) at 4ºC overnight.
2. After dialysis, adjust the protein concentration to 15-30 mM.
3. Add 15 ml of 20 mM FeBABE DMSO solution to 1 ml of the protein solution and incubate it at 37ºC for 1 hour. The final concentration of FeBABE is 0.3 mM (10-20X excess to the protein).
4. Dialyze the reaction mixture in protein storage buffer (10-20 mM Tris, 0.1-0.2 M KCl, 10 mM MgCl2, 0.1 mM EDTA, 50% glycerol, pH 7.6) at 4ºC overnight.

References
Amrita Kumar and Charles P. Moran, Jr., Promoter Activation by Repositioning of RNA Polymerase, JOURNAL OF BACTERIOLOGY, 2008, 190, 3110.

Erin L. Benanti1 and Peter T. Chivers, Helicobacter pylori NikR Protein Exhibits Distinct Conformations When Bound to Different Promoters, THE JOURNAL OF BIOLOGICAL CHEMISTRY, 2011, 286, 15728.

Chih-Chien Wu, Yu-Chun Lin, and Hung-Ta Chen, The TFIIF-Like Rpc37/53 Dimer Lies at the Center of a Protein Network To Connect TFIIIC, Bdp1, and the RNA Polymerase III Active Center, MOLECULAR AND CELLULAR BIOLOGY, 2011, 31, 2715.

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