 |
|
eNOS Antibody Staining Protocol for Immunohistochemistry
Description: Nitric oxide synthase NOS oxidizes a guanidine nitrogen of arginine releasing nitric oxide in the form of a free radical and citrulline. Nitric oxide thus generated acts as a messenger in diverse functions including vasodilation neurotransmission, anti tumor and anti pathogenic activities. NOS is classified under three types: neuronal NOS (nNOS) or brain NOS (bNOS); inducible NOS (iNOS) or macrophage NOS (mNOS); and endothelial NOS (eNOS).
eNOS is a calcium/calmodulin dependent enzyme which undergoes several post translational modifications, including acylation with myristate and palmitate, and phosphorylation on numerous residues. As with the other members of the NOS family, eNOS derives the diffusible multifunctional second messenger NO from L arginine through a series of reactions in which L citrulline is an intermediate. eNOS plays an important role in controlling vascular tone, platelet aggregation, and cardiac myocyte function.
Primary Antibody
|
Name: eNOS Antibody, prediluted |
|
Clone: Rabbit polyclonal |
|
Supplier: Abcam |
|
Catalog Number: ab15280 |
|
Dilution: prediluted ready to use |
|
Incubation Time/Temp: 60 minutes/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 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 of endothelial cells |
| Images: Search image |
Additional Information:
| Species reactivity: Human, mouse, rat, cow, dog and pig |
| Fixation: Formalin fixed paraffin sections |
| Positive Control: HUVEC cells, placenta, skin |
| Negative Control: Omit primary antibody, isotype control or 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 |
Notes:
Not tested on frozen sections |
References: