Description: HNE is a major product of endogenous lipid peroxidation. The w-6-family of polyunsaturated fatty acides may produce HNE as a result of free radical attack. HNE is a highly reactive compound and it can react with several functional groups on biological material, particularly sulfydryl groups to form thioester adduct and then hemiacetals. HNE may also react with histidine and lysine residues of proteins to form stable Michael addition-type of adducts. HNE-modified proteins may display an altered biological functions. Antibodies to HNE will help to visualize the HNE-adducts.
Primary Antibody
Name: HNE Antibody |
Clone: Rabbit anti-Rat |
Supplier: Alpha Diagnostic |
Catalog Number: HNE11-S |
Dilution: 1:500 using IHC-TekTM Antibody Diluent (Cat# IW-1000 or IW-1001) to reduce background and unspecific staining and serum blocking step is NOT needed. |
Incubation Time/Temp: 60 minutes/room temperature |
Device: IHC-TekTM Epitope Retrieval Steamer Set (Cat# IW-1102) |
Buffer/pH value: IHC-TekTM Epitope Retrieval Solution (Cat# IW-1100) |
Heat/Cool Temperature: 95 ºC - 100 ºC/room temperature |
Heat/Cool Time: 20 minutes/20 minutes |
Standard Method: ABC Method or LSAB Method |
Enhanced Method: Polymeric Methods |
Reagent: DAB |
Incubation Time/Temperature: 1-3 minutes/room temperature |
Reagent: Mayer's Hematoxylin |
Staining Time: 30 seconds |
Staining Pattern: Cytoplasmic |
Images: Search image |
Tissue Type: Liver |
Fixation: Formalin fixed paraffin sections |
Positive Control: Liver |
Negative Control: Omit primary antibody, isotype control or absorption control |
Blocking: 2-5% normal serum to reduce unspecific background staining; 0.5-3% H2O2 to block endogenous peroxidase activity; avidin/biotin to block endogenous biotin activity if necessary |
Not tested on frozen sections. |
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