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Co-immunoprecipitations, pull-down assays, far-Western analysis and label transfer reactions are excellent and widely used in vitro approaches to discover or confirm protein interactions.
Finally, the resulting peptide pattern, when analyzed by electrophoresis, immunoblotting, sequencing or mass spectrometry techniques, can provide information relating to the region of contact within the interacting complex. Two purified proteins (bait and prey), preferably of known sequence, that are confirmed or putative binding partners. Additionally, sequence-specific antibodies against the N or C terminal region of the prey will enable detection of the resulting peptides.
To map the interaction site, the cleavage pattern is compared with the unbound end-labeled target protein and known mobility standards.
Bait protein flexibility – use with bait proteins containing endogenous -SH groups, expressed with -SH groups inserted using site-directed mutagenesis, having surface amine groups that can be converted to -SH groups with modification reagents such as 2-iminothiolane. We are Manufacturer and always supplying EDTA-Ferric sodium (CAS:15708-41-5) with competitive price,high quality and regular basis. We have established a privileged position in the industry as manufacturer, supplier and exporter of Tetrasodium EDTA. Dear User, Please use the drop downs below to locate your city by first selecting the country and then the state. These methods verify that an interaction may exist in vivo between the binding partners identified. FeBABE is a sulfhydryl-reactive reagent that incorporates an Fe(III) EDTA moiety into a purified bait protein. Sulfhydryl (-SH) groups may occur naturally as native cysteine residues, or can be introduced by site-directed mutagenesis or by conversion of ε-NH2 groups of lysine with 2-iminothiolane.
Upon establishment of the binding interaction, the Fe(III) chelate (cutting) moiety is activated, causing peptide bonds within reach (12 A) of the labeled bait protein sites to be cleaved.
Several label-detection options, such as radiolabeling, fluorophore labeling or expression tagging, are available to the user.

When expression tagging is used, antibodies against the tag impart the ability to detect the resulting peptides in a Western blot analysis. Mapping the promoter DNA sites proximal to conserved regions of a σ70 in an Escherichia coli RNA polymerase-lacUV5 open promoter complex.
We doesn't provide edta chemical name products or service, please contact them directly and verify their companies info carefully. This acid is a derivative of Ethylenediamine Tetra acetic Acid and is carefully formulated using best-grade chemicals under the strict guidance of our team of experts and at our sound infrastructure setup. If you still can not find your city in the list, please CLICK HERE to submit a request for inclusion of your city. Or complete and post your buying offer of Iron(III)-edta sodium CAS number 21626-24-4 above with . The FeBABE artificial protease reagent and Pierce Protein Interaction Mapping Kit enable more detailed characterization of protein interaction sites. The bromoacetyl group allows the reagent to be covalently attached to proteins at sulfhydryl groups. In the presence of ascorbate and peroxide, the Fe3+ EDTA portion of the reagent is reduced to Fe2+. Proximity mapping the surface of a membrane protein using an artificial protease: Demonstration that the quinone-binding domain of subunit I is near the N-terminal region of subunit II of cytochrome bd. Binding of the σ70 protein to the core subunits of Eschericia coli RNA polymerase studied by iron-EDTA protein footprinting. Synthesis of the protein cutting reagent iron (S)-1-(p-bromoacetamido-benzyl)ethylenediaminetetraacetate and conjugation to cysteine side chains. Our clients can avail this Tetrasodium EDTA in different packaging options from us and at highly reasonable prices.
The kit assembles the FeBABE reagent and other essential reagents and buffers to bring success to both first time and experienced users of this technology.

When activated by ascorbate and peroxide, the Fe(III) EDTA group will cleave peptide bonds, acting as an artificial protease. This reduction promotes the cleavage of peptide bonds by the bait protein in a non-sequence specific manner. Comparison of FeBABE cleavage of the interaction site with cleavage standards and negative reaction controls. The user provides the purified bait protein, a labeled prey protein and a prey-detecting antibody. The outcome of this method is a map (3-D, if the tertiary structure is known) of residues in or near the site of interaction on the target protein.
Reaction mixture containing target protein, FeBABE-cutting protein conjugate and activators (ascorbate and peroxide). Reaction mixture containing target protein and FeBABE-cutting protein conjugate without activators (negative control). Reaction mixture containing target protein, unconjugated cutting protein (no FeBABE) and activators (negative control). EDTA grabs metallic cation such as Lead or Calcium from the process and forms a stable compound that is then excreted from the system. The stability of this bond is vital to get the success in removing the inorganic impurities out of the system.
It is used for removing unwanted inorganic impurities present in the system, which helps in getting superior performance & cost saving.

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