TRAIL Antibody Staining Protocol for Immunohistochemistry


Description: Proteins belonging to the tumor necrosis factor (TNF) superfamily are potent mediators of inflammation and of the immune system. Members of the TNF superfamily include TNFβ, lymphotoxin β (LTβ), CD40L, CD30L, CD27L, Ox40L, 4-1BBL and FAS-L (APO-1). Most TNF family members are type II transmembrane proteins that are proteolytically processed at their carboxy-terminal extracellular domain to form a soluble homotrimeric molecule. The extracellular domain of an additional TNF family member, designated TNF-related apoptosis-inducing ligand (TRAIL) or APO-2L, exhibits 14-28% homology with other members of the TNF family. Like soluble FAS-L, soluble TRAIL will induce apoptosis. The morphological and cellular changes caused by the introduction of soluble TRAIL to Jurkat cells are indistinguishable from those caused by the introduction of soluble FAS-L. Unlike FAS-L, whose expression is more or less restricted to activated T cells, significant levels of TRAIL are observed in many tissues and it is constitutively expressed by some cell lines.


Primary Antibody

Name: Goat Anti-Human TRAIL Antibody

Clone: Goat polyclonal

Supplier: Santa Cruz Biotechnology

Catalog Number: sc-6079

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: Membrane/cytoplasmic
Images: Search image

Additional Information:
Species Reactivity: Human
Fixation: Formalin fixed paraffin sections
Positive Control: Normal breast, liver.
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. if IHC-TekTM Antibody Diluent (Cat# IW-1000 or IW-1001) is used, there is no serum blocking step needed.