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ABN249 Anti-HNE Michael Adducts Antibody, Reduced

Anti-HNE Michael Adducts Antibody, Reduced, Cat. No. ABN249, is a highly specific rabbit polyclonal antibody that targets the reduced form of 4-Hydroxy-2-noneal Michael adducts and has been tested in Western Blotting.

Anti-HNE Michael Adducts Antibody, Reduced MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
Research Category: Neuroscience
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ABN249
100 µL  
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      Overview

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      Key Spec Table

      Species ReactivityKey ApplicationsHostFormatAntibody Type
      B, HWBRbSerumPolyclonal Antibody
      Description
      Catalogue NumberABN249
      ReplacesCalbiochem 393207
      DescriptionAnti-HNE Michael Adducts Antibody, Reduced
      Alternate Names
      • Reduced 4-hydroxy-trans-2-nonenal Michael adducts
      • Reduced HNE Michael adducts
      Background InformationReactive aldehydes generated during lipid peroxidation contribute to cell and tissue damage associated with oxidative stress. Alpha,beta-unsaturated carbonyl compounds such as 4-hydroxy-2-nonenal (HNE), 4-oxo-2-nonenal (ONE), and acrolein, generated from oxidation of polyunsaturated fatty acids (PUFAs), readily interact with nucleophilic groups on proteins, leading to intramolecular or intermolecular protein crosslinking. Two types of HNE modifications exist, the first type is formed by a 1:1 Michael adduct reaction with lysine amine, histine imidazole, or cysteine sulfhydryl, while the second type is produced by one HNE reacting simultaneously with two free amine groups from two N-alpha-acetyllysines (NALs), so-called Michael-Schiff base adduct. In the latter case, the initial nonfluorescent 2:1 lysine-HNE Michael adduct-Schiff base further undergoes oxidative cyclization to form a fluorescent 2-hydroxy-3-imino-1,2-dihydropyrrol derivative commonly referred to as HNE fluorophore. Reversibly formed HNE-Lys Michael adduct is not stable and reduction by sodium borohydride (NaBH4) is often employed to preserve Lys HNE mofidication in protein samples. Reductive stabilization by NaBH4 treatment is also used to prevent 4-HNE loss from 4-HNE-Cys Michael adduct by retro-Michael addition reaction (RMA) during collision-induced dissociation (CID) and electron transfer dissociation (ETD) MS/MS.
      References
      Product Information
      FormatSerum
      Control
      • HNE-modifed BSA chemically reduced by sodium borohydride.
      PresentationRabbit polyclonal antibody serum with 0.05% sodium azide.
      Quality LevelMQ100
      Applications
      ApplicationAnti-HNE Michael Adducts Antibody, Reduced, Cat. No. ABN249, is a highly specific rabbit polyclonal antibody that targets the reduced form of 4-Hydroxy-2-noneal Michael adducts and has been tested in Western Blotting.
      Key Applications
      • Western Blotting
      Application NotesApplication Notes:
      1. ABN249 replaces Calbiochem 393207. ABN249 is expected to work in immunocytochemistry and immunohistochemistry. Please refer to the following 393207 citations as examples of sample preparations and detection in these two types of applications:

      Usatyuk, P.V., et al. (2006). J. Biol. Chem. 281(46):35554-35566 (Immunocytochemistry).
      McCormack, A.L., et al. (2005). J. Neurochem. 93(4):1030-1037 (Immunohistochemistry).

      2. Anti-HNE Fluorophore, Cat. No. 393206, is also available for the detection of HNE Fluorophore by ELISA, Immunohistochemistry, and Western blotting.

      3. To generate a positive control for Western blotting, treat a 2 mg/mL BSA solution in 10 mM KH2PO4, pH 7.4, with 0.1 to 0.5 mM HNE and incubate for 1 hr at 37°C. The reaction can be monitored by the decrease of HNE absorbance at 224 nm (extinction coefficient 13,750/M). Free HNE can be removed by gel filtration and dialysis following the chemical reaction.

      HNE Michael adducts in protein solutions must be chemically reduced by sodium borohydride (NaBH4) prior to analysis using ABN1348. A freshly prepared 100 mM NaBH4 solution in 1 mM NaOH is added 1:9 (V:V) to the protein sample solution to a final NaBH4 concentration of 10 mM. The protein solution is kept at room temperature for 10 to 30 min prior to quenching with the addition of 5 to 10 molar equivalents of acetone.
      Biological Information
      ImmunogenHNE-modifed KLH chemically reduced by sodium borohydride.
      ConcentrationPlease refer to lot specific datasheet.
      HostRabbit
      SpecificityThis polyclonal antiserum detected HNE-modified BSA only after, but not before, reduction by sodium borohydride.
      Species Reactivity
      • Bovine
      • Human
      Species Reactivity NoteTarget modification is not species-specific.
      Antibody TypePolyclonal Antibody
      Purification MethodUnpurified.
      Molecular WeightVariable depending on the size(s) of the HNE-modified protein(s). Uncharacterized bands may be observed in some lysate(s).
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Quality AssuranceEvaluated by Western Blotting in Reduced HNE-BSA.

      Western Blotting Analysis: A 1:10,000 of this antibody detected 10 ng of HNE-modified BSA only after, but not before, reduction by sodium borohydride.
      Usage Statement
      • Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
      Storage and Shipping Information
      Storage ConditionsStable for 1 year at -20°C from date of receipt.

      Handling Recommendations: Upon receipt and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.
      Packaging Information
      Material Size100 µL
      Transport Information
      Supplemental Information
      Specifications
      Global Trade Item Number
      Catalogue Number GTIN
      ABN249 04054839057182

      Documentation

      Anti-HNE Michael Adducts Antibody, Reduced SDS

      Title

      Safety Data Sheet (SDS) 

      Anti-HNE Michael Adducts Antibody, Reduced Certificates of Analysis

      TitleLot Number
      Anti-HNE Michael Adducts, Reduced - 3393273 3393273
      Anti-HNE Michael Adducts, Reduced - 3800055 3800055
      Anti-HNE Michael Adducts, Reduced - 4002928 4002928
      Anti-HNE Michael Adducts, Reduced - 4157290 4157290
      Anti-HNE Michael Adducts, Reduced -Q2737541 Q2737541
      Anti-HNE Michael Adducts, Reduced Polyclonal Antibody 3133543