203110 Biotin-XX-Hydrazide

203110
  
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      Overview

      Replacement Information

      Key Spec Table

      Description
      Overview

      This product has been discontinued.





      Biotinylates oxidized sugars on glycoproteins and inserts a 13-atom spacer between biotin and the target ligand, thereby alleviating steric hindrance.
      Catalogue Number203110
      Brand Family Calbiochem®
      References
      ReferencesLeary, J.J., et al. 1983. Proc. Natl. Acad. Sci. USA 80, 4045.
      Hoffman, K., et al. 1982. Biochemistry 21, 978.
      Bayer, E.A., et al. 1978. Methods Enzymol. 62, 308.
      Product Information
      CAS number211237-33-1
      FormWhite solid
      Structure formula ImageStructure formula Image
      Applications
      Biological Information
      Purity>95% by TLC
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Storage and Shipping Information
      Ship Code Ambient Temperature Only
      Toxicity Standard Handling
      Storage +2°C to +8°C
      Do not freeze Ok to freeze
      Special InstructionsFollowing reconstitution, aliquot and freeze (-20°C). Stock solutions are stable for up to 1 month at -20°C.
      Packaging Information
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      Certificates of Analysis

      TitleLot Number
      203110

      References

      Reference overview
      Leary, J.J., et al. 1983. Proc. Natl. Acad. Sci. USA 80, 4045.
      Hoffman, K., et al. 1982. Biochemistry 21, 978.
      Bayer, E.A., et al. 1978. Methods Enzymol. 62, 308.

      Brochure

      Title
      Protein Modifications Tools

      Citations

      Title
    • Maryse Delehedde, et al. (2002) Fibroblast growth factor-2 binds to small heparin-derived oligosaccharides and stimulates a sustained phosphorylation of p42/44 mitogen-activated protein kinase and proliferation of rat mammary fibroblasts. Biochemistry Journal 366, 235-244.
    • Data Sheet

      Note that this data sheet is not lot-specific and is representative of the current specifications for this product. Please consult the vial label and the certificate of analysis for information on specific lots. Also note that shipping conditions may differ from storage conditions.

      Revision31-March-2011 RFH
      DescriptionBiotinylates oxidized sugars on glycoproteins and inserts a 13-atom spacer between biotin and the target ligand, thereby alleviating steric hindrance. Biotin Hydrazide (BHZ) is an extremely valuable biotinylating reagent. In general, BHZ reacts covalently with oxidation-induced aldehydes of membrane bound proteins and/or glycoproteins. BHZ is commonly employed as an alternative to biotin-N-hydroxysuccinimide ester (biotin-NHS) in the labeling of immunoglobulins. Biotin-NHS reacts predominantly with the ε-amino groups of lysine side chains. Although the use of biotin-NHS is common, modification of amino acid residues within or in close proximity to the antigen binding site may result in the loss of immunological activity. BHZ avoids this by reacting with carbohydrate moieties of immunoglobulins. BHZ is also widely used as a direct label of cell membrane proteins.
      FormWhite solid
      Recommended reaction conditions
      Biotin-XX-Hydrazide Protocol There are several different protocols to generate the reactive aldehydes necessary for labeling with BHZ, using either chemical or enzymatic methods. Sialyl groups can be directly oxidized to a reactive aldehyde group with sodium periodate. Sialyl groups can also be removed by neuraminidase and the penultimate galactose residue is then oxidized by galactose oxidase. N-Acetylgalactosaminyl residues or terminal galactosyl residues can also be directly oxidized using galactose oxidase. Vicinal hydroxyl groups of other sugars can be periodate oxidized.
      Chemical or enzymatic oxidation of aldehydes is not the only procedure that can be used with BHZ. A novel one step procedure was developed to introduce biotin into nucleic acids for non-isotropic hybridization. Bisulfite was used to catalyze the reaction between BHZ and unpaired cytosine residues in single stranded DNA. In another published procedure, BHZ was coupled to surface expressed proteins through a very reactive nitrene intermediate generated by the reaction of sodium nitrate on BHZ. An alternative method was used to modify β-nerve growth factor via a carboxyl group substitution using BHZ and the coupling reagent 1-ethyl-3-(dimethylaminopropyl)-carbodiimide.

      The following is a general protocol for labeling with BHZ. Sodium periodate is used to oxidize both glycoproteins and cell membrane proteins, whereas the enzymatic method is used primarily with cell membrane labeling. Only the periodate method is presented here.

      General Labeling of Glycoproteins:
      1. Oxidize 1-2 mg/ml of protein in 100 mM sodium acetate buffer, pH 5.5, containing 10 mM sodium periodate and incubate for 30 min at 0°C.
      2. Quench excess periodate by the addition of 15 mM glycerol or 20 mM sodium sulfite for 5 min at 0°C.
      3. Dialyze the protein solution overnight against 100 mM sodium acetate, pH 5.5, at 4-20°C.
      4. Add 1-10 mM Biotin-XX-Hydrazide and incubate for 1-2 h at 20°C.
      5. Dialyze against PBS overnight at 4-20°C.

      General Labeling of Cell Membrane Proteins:
      1. A given cell suspension (107) is treated with 1 mM sodium periodate in PBS for 30 min in an ice bath.
      2. Centrifuge the cells, wash 3X with PBS, and re-suspend in PBS containing 10 mM Biotin-Hydrazide.
      3. Incubate for 30 min at 20°C. Addition of 1 mM MnCl2 enhances binding of BHZ to aldehydes.
      4. Centrifuge the cells, wash 3X with PBS, and re-suspend in an appropriate buffer to the desired volume.*
      *Some references treat the BHZ modified cells with 1 mM sodium borohydride to increase stability of the linkage, yet others say it is not necessary.
      CAS number211237-33-1
      Structure formulaStructure formula
      Purity>95% by TLC
      SolubilityDMF (10 mg/ml) or DMSO (10 mg/ml)
      Storage +2°C to +8°C
      Do Not Freeze Ok to freeze
      Special InstructionsFollowing reconstitution, aliquot and freeze (-20°C). Stock solutions are stable for up to 1 month at -20°C.
      Toxicity Standard Handling
      ReferencesLeary, J.J., et al. 1983. Proc. Natl. Acad. Sci. USA 80, 4045.
      Hoffman, K., et al. 1982. Biochemistry 21, 978.
      Bayer, E.A., et al. 1978. Methods Enzymol. 62, 308.
      Citation
    • Maryse Delehedde, et al. (2002) Fibroblast growth factor-2 binds to small heparin-derived oligosaccharides and stimulates a sustained phosphorylation of p42/44 mitogen-activated protein kinase and proliferation of rat mammary fibroblasts. Biochemistry Journal 366, 235-244.