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DR4 Antibody Staining Protocol for Immunohistochemistry
Description: Tumor necrosis factor (TNF) is a pleiotropic cytokine whose function is mediated by two distinct cell surface receptors, designated TNF-R1 and TNF-R2, which are expressed on most cell types. TNF function is primarily mediated through TNF-R1 signaling. Both receptors belong to the growing TNF receptor superfamily which, includes FAS antigen and CD40. TNF-R1 contains a cytoplasmic motif, termed the “death domain,” that has been found to be necessary for the transduction of the apoptotic signal. The death domain is also found in several other receptors, including FAS, DR2 (or TRUNDD), DR3 (Death Receptor 3), DR4 and DR5. TRUNDD, DR4 and DR5 are receptors for the apoptosis-inducing cytokine TRAIL. A non-death domain-containing receptor, designated decoy receptor (DcR1 or TRID), also specifically associates with TRAIL and may play a role in cellular resistance to apoptotic stimuli.
Primary Antibody
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Name: Goat Anti-Human DR4 Antibody |
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Clone: Goat polyclonal |
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Supplier: Santa Cruz Biotechnology |
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Catalog Number: sc-6823 |
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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. |
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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: Cytoplasmic |
| Images: Search image |
Additional Information:
| Species Reactivity: Human |
| Fixation: Formalin fixed paraffin sections |
| Positive Control: Normal breast |
| 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: