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

Name: Thyroid Transcription Factor, TTF-1 Antibody

Clone: 8G7G31, Mouse monoclonal

Supplier: Dako

Catalog Number: M3575

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: 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: 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

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.