Anti-Phospho-eNOS (Thr495) Polyclonal Antibody 2 is a Rabbit antibody targeting Phospho-eNOS (Thr495). Anti-Phospho-eNOS (Thr495) Polyclonal Antibody 2 can be used in WB,FCM.

IgG

Human,Mouse (predicted:Rat,Dog,Pig,Cow,Rabbit,Sheep,GuineaPig)

1. Sample: k562 (human) Cell Lysate at 30 μg
Primary: Anti-Phospho-eNOS (Thr495) (Ser319) (TMAB-10649) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 133 kD
Observed band size: 133 kD
2. Sample:
Lane1: Liver (Mouse) Lysate at 30 μg
Lane2: Kidney (Mouse) Cell Lysate at 30 μg
Primary: Anti-Phospho-eNOS (TMAB-10649) at 1:300 dilution;
Secondary: HRP conjugated Goat-Anti-Rabbit IgGat 1: 3000 dilution;
Predicted band size: 133 kD
Observed band size: 135 kD
3. Blank control: U937.
Primary Antibody (green line): Rabbit Anti-Phospho-eNOS (Thr495) antibody (TMAB-10649)
Dilution: 2 μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody: Goat anti-rabbit IgG-PE
Dilution: 1 μg /test.
Protocol
The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 0.1% PBST for 20 min at room temperature. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.

WB: 1:500-2000; FCM: 2μg/Test

Polyclonal

KLH conjugated Synthesised phosphopeptide: human eNOS around the phosphorylation site of Thr495

Human

NOS3

Nitric oxide synthase, endothelial

Produces nitric oxide (NO) which is implicated in vascular smooth muscle relaxation through a cGMP-mediated signal transduction pathway. NO mediates vascular endothelial growth factor (VEGF)-induced angiogenesis in coronary vessels and promotes blood clotting through the activation of platelets.
Isoform eNOS13C: Lacks eNOS activity, dominant-negative form that may down-regulate eNOS activity by forming heterodimers with isoform 1.