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Neutrophil Elastase Antibody Staining Protocol for Immunohistochemistry
Description: Monoclonal Mouse Anti-Human Neutrophil Elastase, Clone NP57, is intended for use in immunocytochemistry. The antibody labels neutrophils and neutrophil precursors, and also reacts (although more weakly) with a minor population of blood monocytes. Neoplastic cells in about 70% of acute myeloid leukaemias are labelled by the antibody, and it is a useful supplement to other antibodies to myeloid markers in the differential identification of acute myeloid leukaemias and extramedullary myeloid cell tumours. Differential identification is aided by the results from a panel of antibodies and traditional histological and enzyme cytochemical methods. Interpretation must be made within the context of the patient’s clinical history and other diagnostic tests by a qualified pathologist.
Primary Antibody
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Name: Neutrophil Elastase Antibody |
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Clone: NP57, Mouse anti-Human |
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Supplier: Dako |
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Catalog Number: M0752 |
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Dilution: 1:100 using IHC-TekTM Antibody Diluent (Cat# IW-1000 or IW-1001) to reduce background and unspecific staining and serum blocking step is NOT needed. |
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Incubation Time/Temp: 60 min/room temperature |
Antigen Retrieval
| Device: No pretreatment needed. Proteinase K pretreatment may increase staining intensity. |
| Buffer/pH value: IHC-TekTM Epitope Retrieval Solution (Cat# IW-1100) |
| Heat/Cool Temperature: 37 ºC/room temperature |
| Heat/Cool Time: 20 minutes/20 minutes |
Detection Methods
| Standard Method: ABC Method or LSAB Method |
| Enhanced Method: Polymeric Methods |
Chromogen Substrate
| Reagent: DAB |
| Incubation Time/Temperature: 1-3 minutes/room temperature |
Counterstain
| Reagent: Mayer's Hematoxylin |
| Staining Time: 30 seconds |
Results:
| Staining Pattern: Cytoplasmic |
| Images: Search image |
Additional Information:
| Species Reactivity: Human |
| Fixation: Formalin fixed paraffin sections, acetone fixed frozen sections, or cell smears |
| Positive Control: Tonsil, bone marrow |
| Negative Control: Omit primary antibody, isotype control, 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 |
References: