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IL-1ra (IL-1 receptor antagonist) Antibody Staining Protocol for Immunohistochemistry
Description: Two forms of interleukin-1, designated IL-1α and IL-1β, have been described. Although encoded by distinct genes and exhibiting roughly only 25% sequence identity, IL-1α and IL-1β bind to the same receptor and seem to elicit similar biological responses. Both proteins are synthesized as 31 kDa precursors that are processed to 17 kDa mature polypeptides. IL-1 production is generally thought to be associated with inflammation, but it has also been shown to be expressed during kidney development, thymocyte differentiation and cartilage degradation. IL-1 plays a critical role in the regulation of immune response and inflammation acting as an activator of T and B lymphocytes and natural killer (NK) cells. In T cells, IL-1 stimulates the production of IL-2 and selectively inhibits IL-4 expression. IL-1 receptor antagonist (IL-1ra) is a cytokine that inhibits IL-1α and IL-1β binding to interleukin receptors.
Primary Antibody
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Name: IL-1ra (H-110) Antibody |
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Clone: Rabbit polyclonal |
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Supplier: Santa Cruz Biotechnology |
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Catalog Number: sc-25444 |
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Dilution: 1:100 - 1:200 depending on detection system and tissue species 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: Gill's Hematoxylin or Mayer's Hematoxylin |
| Staining Time: 30 seconds |
Results:
| Staining Pattern: Cytoplasmic of fibroblasts, endothelial cells, extracellular matrix |
| Images: Search image |
Additional Information:
| Species Reactivity: Human, mouse, rat, guinea pig |
| Fixation: Formalin fixed paraffin sections |
| Positive Control: Skin, tonsil, kidney |
| 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: