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IL-6 Antibody Staining Protocol for Immunohistochemistry
Description: Interleukin 6 (IL6) is a multifunctional 26 kD protein originally discovered in the medium of RNA-stimulated fibroblastoid cells. IL6 appears to be directly involved in the responses that occur after infection and cellular injury, and it may prove to be as important as IL1 and TNF-a in regulating the acute phaseresponse. IL6 is reported to be produced by fibroblasts, activated T cells, activated monocytes or macrophages and endothelial cells. It acts upon a variety of cells including fibroblasts, myeloid progenitor cells, T cells, B cells and hepatocytes. In addition, IL6 appears to interact with IL2 in the proliferation of T lymphocytes. IL6 potentiates the proliferative effect of IL3 on multipotential hematopoietic progenitors.
Primary Antibody
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Name: IL-6 Antibody |
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Clone: Rabbit anti-Human |
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Supplier: Abcam |
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Catalog Number: ab6672 |
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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: Incubator |
| Buffer/pH value: IHC-TekTM Proteinase K Solution (Cat# IW-1101) |
| Heat/Cool Temperature: 37 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: Secreted by fibroblasts, macrophages and endothelial cells, etc. |
| Images: Search image |
Additional Information:
| Tissue Type: Skin |
| Fixation: Formalin fixed paraffin sections |
| Positive Control: Skin |
| 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: