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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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Key Specifications 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
Catalog 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