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Angiotensin Converting Enzyme 1 (ACE-1) Antibody Staining Protocol for Immunohistochemistry
Description: ACE converts angiotensin I to angiotensin II by release of the terminal His-Leu, this results in an increase of the vasoconstrictor activity of angiotensin. Also able to inactivate bradykinin, a potent vasodilatator. ACE exists in two forms, a 170KD somatic form and a 90KD germinal form. The somatic form is expressed by endothelial cells (especially those of lung capillaries and arterioles), epithelial cells (especially in proximal renal tubules and small intestine), by some neuronal cells and variably by some macrophages and T lymphocytes. The germinal form is expressed by spermatozoa.
Primary Antibody
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Name: Mouse Anti-ACE-1 Antibody |
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Clone: Mouse monoclonal (2E2) |
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Supplier: Abcam |
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Catalog Number: ab11734 |
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Dilution: 1:10 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 Chromogen Substrate |
| Incubation Time/Temperature: 1-3 minutes/room temperature |
Counterstain
| Reagent: Mayer's Hematoxylin |
| Staining Time: 30 seconds |
Results:
| Staining Pattern: Cell membrane |
| Images: Search image |
Additional Information:
| Species Reactivity: Mouse, Rat, Rabbit, Guinea pig, Cow, Cat, Dog, Human, Monkey |
| Fixation: Formalin fixed paraffin sections |
| Positive Control: Lung, 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: