Immunofluorescence analysis of eNOS was done on 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with eNOS Rabbit Polyclonal Antibody (PA516887) at 2ug/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d is a merged image showing cytoplasmic localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Bovine, Dog, Human, Mouse, Pig, Rat|
|Host / Isotype||Rabbit / IgG|
|Immunogen||A synthetic peptide derived from C-ternimal of human eNOS|
|Storage buffer||PBS, pH 7.6, with 0.2% BSA|
|Contains||15mM sodium azide|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|Immunohistochemistry (Paraffin) (IHC (P))||1:20|
|Western Blot (WB)||1:50-1:500|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
PA5-16887 targets eNOS in immunohistochemistry (paraffin) applications and shows reactivity with Bovine, Canine, Human, mouse, Porcine, and Rat samples. Suggested positive control is capillary endothelium and regenerative endothelium.
The PA5-16887 immunogen is a synthetic peptide derived from C-ternimal of human eNOS.
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); induciblr NOS (iNOS) or macrophage NOS (mNOS); and endothelial NOS (eNOS). This antibody reacts with eNOS.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.