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

Name: Neutrophil Elastase Antibody

Clone: NP57, Mouse anti-Human

Supplier: Dako

Catalog Number: M0752

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.

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

Reagent: Mayer's Hematoxylin
Staining Time: 30 seconds

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