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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
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Name: Goat Anti-Human TRAIL Antibody |
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Clone: Goat polyclonal |
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Supplier: Santa Cruz Biotechnology |
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Catalog Number: sc-6079 |
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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: 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. |
References: