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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
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Name: Mast Cell Tryptase Antibody |
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Clone: AA1, Mouse anti-Human |
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Supplier: Dako |
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Catalog Number: M7052 |
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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. |
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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 |
Counterstain
| Reagent: Mayer's Hematoxylin |
| Staining Time: 30 seconds |
Results:
| 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 |
References: