TGF beta RI Antibody Staining Protocol for Immunohistochemistry


Description: A total of three members of the TGF ß family, TGFß1, TGFß2 and TGFß3, have been identified in mammals. Each is synthesized as a latent precursor that is subsequently cleaved forming the 112 amino acid growth factor which becomes active upon dimerization. TGFßs mediate their activity by high affinity binding to the type II receptor 70 kDa transmembrane protein with a cytoplasmic serine-threonine kinase domain. For signaling growth inhibition and early gene responses the type II receptor requires both its kinase activity and association with a 53 kDa TGFß-binding protein, designated the type I receptor. Two independent groups have recently described the cloning and sequence analysis of genes encoding 53 kDa TGFß type I receptor proteins (94 kDa receptor complex) designated ALK-5 (TßR-1) and TSR-1, respectively.


Primary Antibody

Name: TGF beta RI (V-22) Antibody

Clone: Rabbit polyclonal

Supplier: Santa Cruz Biotechnology

Catalog Number: sc-398

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:
Species Reactivity: Human, mouse, rat
Fixation: Formalin fixed paraffin sections
Positive Control: Prostate
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