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TNF-alpha Antibody Staining Protocol for Immunohistochemistry
Description: Tumor necrosis factor-alpha (TNFA) also known as TNFR1, It is a potent cytokine, elicits a broad spectrum of biologic responses that are mediated by binding to a cell surface receptor. There are 2 different proteins that serve as major receptors for TNF-alpha, one associated with myeloid cells and one associated with epithelial cells. TNFR1 maps to 12p13. TNFR1 signaling is also known to activate the transcription factor NF-kappa B and promote survival.
Primary Antibody
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Name: TNF-alpha IHC Antibody |
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Clone: Rabbit polyclonal |
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Supplier: IHC World |
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Catalog Number: IW-PA1079 |
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Dilution: Ready to Use |
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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: Gill's Hematoxylin or Mayer's Hematoxylin |
| Staining Time: 30 seconds |
Results:
| Staining Pattern: Cytoplasmic |
| Images: Search image |
Additional Information:
| Species Reactivity: Human, mouse, rat, rabbit. |
| Fixation: Formalin fixed paraffin sections |
| Positive Control: Human mammary cancer |
| 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 |
References: