KLH conjugated Synthesised phosphopeptide derived from human eNOS around the phosphorylation site of Thr495:KK(p-T)FK
Format：
Liquid
Storage instructions：
Shipped at 4℃. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles.
Concentration：
1mg/ml
Clonality：
Polyclonal
Isotype：
IgG
Applications：
WB=1:500-2000ELISA=1:5000-10000IHC-P=1:100-500IHC-F=1:100-500Flow-Cyt=2ug/TestIF=1:100-500（Paraffin sections need to do antigen repair）not yet tested in other applications.optimal dilutions/concentrations should be determined by the end user.
Host：
Rabbit
Product Overview：
Sample: K562(Human) Cell Lysate at 30 ugPrimary: Anti-Phospho-eNOS (Thr495) (Ser319) (SL3731R) at 1/300 dilutionSecondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilutionPredicted band size: 133 kDObserved band size: 133 kDSample: Lane1: Liver (Mouse) Lysate at 30 ugLane2: Kidney (Mouse) Cell Lysate at 30 ugPrimary: Anti-Phospho-eNOS (SL3731R) at 1:300 dilution; Secondary: HRP conjugated Goat-Anti-Rabbit IgG(bse-0295G) at 1: 3000 dilution; Predicted band size : 133kDObserved band size : 135kDBlank control:U937. Primary Antibody (green line): Rabbit Anti-Phospho-eNOS (Thr495) antibody (SL3731R) Dilution: 2μg /10^6 cells; Isotype Control Antibody (orange line): Rabbit IgG . Secondary Antibody : Goat anti-rabbit IgG-PEDilution: 1μg /test. ProtocolThe cells were fixed with 4% PFA (10min 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.
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.
Function: 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.
Subunit: Homodimer. Interacts with NOSIP and NOSTRIN.
Subcellular Location: Cell membrane. Membrane, caveola. Cytoplasm, cytoskeleton. Golgi apparatus. Note=Specifically associates with actin cytoskeleton in the G2 phase of the cell cycle; which is favored by interaction with NOSIP and results in a reduced enzymatic activity.
Tissue Specificity: Platelets, placenta, liver and kidney.
Post-translational modifications: Phosphorylation by AMPK at Ser-1177 in the presence of Ca(2+)-calmodulin (CaM) activates activity. In absence of Ca(2+)-calmodulin, AMPK also phosphorylates Thr-495, resulting in inhibition of activity. Phosphorylation of Ser-114 by CDK5 reduces activity.
Similarity: Belongs to the NOS family.
Contains 1 FAD-binding FR-type domain.
Contains 1 flavodoxin-like domain.
催化生物体内一氧化氮（NO）生成的酶。分神经型一氧化氮合成的酶（nNOS or NOS-1）、诱导型一氧化氮合成的酶（iNOS or NOS-2）、内皮型一氧化氮合成的酶（eNOS or NOS-3）。此抗体识别分子量为135kDa的内皮型一氧化氮合成的酶。
Picture
Sample:
K562(Human) Cell Lysate at 30 ug
Primary: Anti-Phospho-eNOS (Thr495) (Ser319) (SL3731R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 133 kD
Observed band size: 133 kD
Sample:
Lane1: Liver (Mouse) Lysate at 30 ug
Lane2: Kidney (Mouse) Cell Lysate at 30 ug
Primary: Anti-Phospho-eNOS (SL3731R) at 1:300 dilution;
Secondary: HRP conjugated Goat-Anti-Rabbit IgG(bse-0295G) at 1: 3000 dilution;
Predicted band size : 133kD
Observed band size : 135kD
Blank control:U937.
Primary Antibody (green line): Rabbit Anti-Phospho-eNOS (Thr495) antibody (SL3731R)
Dilution: 2μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody : Goat anti-rabbit IgG-PE
Dilution: 1μg /test.
Protocol
The cells were fixed with 4% PFA (10min 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.
Product Feedback Wall
Specific References (1) | SL3731R has been referenced in 1 publications.
[IF=4.99] Papinska, A. M., et al. "Angiotensin‐(1‐7) administration benefits cardiac, renal, and progenitor cell function in db/db mice." British Journal of Pharmacology(2015). IHSLCP ; Mouse.
