This antibody gave a positive signal in the following Human tissue lysates: Thymus; Pancreas; Small Intestine; Stomach Tumor.
IHC-P: FFPE human lung tissue sections
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pH: 7.40 Preservative: 0.02% Sodium azide Constituent: PBS Note: Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 µg/ml. Detects a band of approximately 30, 32 kDa (predicted molecular weight: 30 kDa).
Use a concentration of 0.1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Mast cells contain a number of preformed chemical mediators such as histamine, chymase, carboxypeptidase and proteolytic tryptase.
Human Mast Cell Tryptase is considered to be an important marker of mast cell activation as well as an important mediator of inflammation.
Secreted. Note: released from the secretory granules upon mast cell activation.
Western blot - Anti-Mast Cell Tryptase antibody (ab64192)
All lanes : Anti-Mast Cell Tryptase antibody (ab64192) at 1 µg/ml
Lane 1 : Human thymus tissue lysate - total protein (ab30146) Lane 2 : Human pancreas tissue lysate - total protein (ab29816) Lane 3 : Human small intestine tissue lysate - total protein (ab29276) Lane 4 : Human stomach tumor tissue lysate (adenocarcinoma) - total protein (ab29678)
Lysates/proteins at 10 µg per lane.
Secondary All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Mast Cell Tryptase contains a number of potential glycosylation sites (SwissProt) which may explain the higher migration pattern observed.
The band observed at 30 kDa could potentially be a cleaved form of Mast Cell Tryptase due to the presence of an 18 amino acid signal peptide.
IHC image of ab64192 staining in human lung formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab64192, 0.1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.