Mast Cell Tryptase Antibody Staining Protocol for Immunohistochemistry


Description: This antibody labels human mast cells and is a useful tool for the recognition of very atypical or immature mast cells (MC) in mast cell leukaemia, and for the detection of small, even minute, dense focal MC infiltrates in staging procedures in patients with known cutaneous mastocytosis.


Primary Antibody

Name: Mast Cell Tryptase Antibody

Clone: AA1, Mouse anti-Human

Supplier: Dako

Catalog Number: M7052

Dilution: 1:200 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: IHC-TekTM Epitope Retrieval Steamer Set (Cat# IW-1102)
Buffer/pH value: IHC-TekTM Epitope Retrieval Solution (Cat# IW-1100)
Heat/Cool Temperature: 95-100 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:
Tissue Type: Bone marrow, skin
Fixation: Formalin fixed paraffin sections
Positive Control: Human tonsil, bone marrow, lung, skin, colon, gastric mucosa.
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