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Thyroid Transcription Factor (TTF-1) Antibody Staining Protocol for Immunohistochemistry
Description: Thyroid transcription factor-1 (TTF-1) belongs to a family of homeodomain transcription factors and and plays a role in regulating genes expressed within the thyroid, lung and brain which include thyroglobulin, thyroid peroxidase, Clara cell secretory protein and surfactant proteins. In addition to its role as a tissue-specific transcriptional activator in adult organs, TTF-1 may also function in organogenesis. Gene targeting studies has shown TTF-1 to be essential for the proper development of the thyroid and lungs. TTF-1 is selectively expressed in thyroid, lung and diencephalon. TTF-1 has been identified as a transcriptional regulator of thyroid-specific genes and has also been shown to be important in the activation of pulmonary-specific differentiation genes. Monoclonal anti-rat TTF-1 clone 8G7G3/1 (anti-TTF-1) has also been shown to react with rat, mouse and human TTF-1.
Primary Antibody
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Name: Thyroid Transcription Factor, TTF-1 Antibody |
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Clone: 8G7G31, Mouse monoclonal |
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Supplier: Dako |
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Catalog Number: M3575 |
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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: 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: Nuclear |
| Images: Search image |
Additional Information:
| Species Reactivity: Human, mouse, rat |
| Fixation: Formalin fixed paraffin sections, or acetone fixed frozen sections (no antigen retrieval needed) |
| Positive Control: Lung, thyroid |
| 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 |
Note:
TTF-1 (Clone SPT24, Novocastra, Cat# NCL-TTF-1) works well at 1:100 dilution with Tris-EDTA Buffer Epitope Retrieval Method. TTF-1 (Clone 8G7G3/1, Labvision, Cat# MS-699-p1) also works well at 1:200 dilution with Citrate Buffer Epitope Retrieval Method.
References: