WB
Western blot analysis of eNOS on different lysates with Rabbit anti-eNOS antibody at 1/5,000 dilution. Lane 1: EA.hy926 cell lysate (20 µg/Lane), Lane 2: HeLa cell lysate (negative) (20 µg/Lane), Lane 3: bEnd.3 cell lysate (20 µg/Lane), Lane 4: RAW264.7 cell lysate (negative) (20 µg/Lane), Lane 5: Mouse placenta tissue lysate (30 µg/Lane), Lane 6: Mouse kidney tissue lysate (30 µg/Lane), Lane 7: Rat placenta tissue lysate (30 µg/Lane), Lane 8: Rat kidney tissue lysate (30 µg/Lane), Exposure time: Lane 1-2: 10 seconds; Lane 3-8: 3 minutes; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/5,000 dilution was used in primary antibody dilution at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.IHC
Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-eNOS antibody at 1/50 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/50 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.IHC
Immunohistochemical analysis of paraffin-embedded mouse spleen tissue with Rabbit anti-eNOS antibody at 1/50 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/50 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.IHC
Immunohistochemical analysis of paraffin-embedded rat spleen tissue with Rabbit anti-eNOS antibody at 1/50 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/50 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.ICC/IF
Immunocytochemistry analysis of EA.hy926 (positive) and HeLa (negative) labeling eNOS with Rabbit anti-eNOS antibody at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-eNOS antibody at 1/50 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.FC
Flow cytometric analysis of HeLa (left, negative) and EA.hy926 (right, positive) cells labeling eNOS. Cells were fixed and permeabilized. Then stained with the primary antibody (1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).IP
eNOS was immunoprecipitated from 0.2 mg bEnd.3 cell lysate with Rabbit anti-eNOS antibody at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate usingRabbit anti-eNOS antibody at 1/5,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: bEnd.3 cell lysate (input), Lane 2: Rabbit anti-eNOS antibody IP in bEnd.3 cell lysate, Lane 3: Rabbit IgG instead of Rabbit anti-eNOS antibody in bEnd.3 cell lysate, Blocking/Dilution buffer: primary antibody dilution, Exposure time: 3 minutes.| Product Name | eNOS Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Recombinant protein within human eNOS aa 950-1,203. |
| Clonality | monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | FCICC/IFIHCIPWB |
| WB:1:5000 IHC:1:50-1:200 ICC/IF:1:50 FC:1:1000 IP:1-2μg/sample |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | NOS3 |
|---|---|
| Gene Synonyms | eNOS ECNOS MYMY8 |
| Gene Full Name | nitric oxide synthase 3 |
| Gene Summary | Nitric oxide is a reactive free radical which acts as a biologic mediator in several processes, including neurotransmission and antimicrobial and antitumoral activities. Nitric oxide is synthesized from L-arginine by nitric oxide synthases. Variations in this gene are associated with susceptibility to coronary spasm. Alternative splicing and the use of alternative promoters results in multiple transcript variants. [provided by RefSeq, Oct 2016] |
| Alternative Names | cNOS antibody Constitutive NOS antibody EC NOS antibody EC-NOS antibody ecNOS antibody Endothelial nitric oxidase synthase antibody Endothelial nitric oxide synthase antibody Endothelial nitric oxide synthase 3 antibody Endothelial NOS antibody eNOS antibody cNOS antibody Constitutive NOS antibody EC NOS antibody EC-NOS antibody ecNOS antibody Endothelial nitric oxidase synthase antibody Endothelial nitric oxide synthase antibody Endothelial nitric oxide synthase 3 antibody Endothelial NOS antibody eNOS antibody Nitric oxide synthase 3 (endothelial cell) antibody Nitric oxide synthase 3 antibody Nitric oxide synthase 3 endothelial cell antibody Nitric oxide synthase endothelial antibody Nitric oxide synthase endothelial antibody NOS 3 antibody NOS III antibody NOS type III antibody NOS3 antibody NOS3_HUMAN antibody NOSIII antibody |
| Molecular Weight(MW) | 133 kDa |
| Cellular Localization | Cell membrane, Membrane, caveola, Cytoplasm, cytoskeleton, Golgi apparatus. |
WB
Western blot analysis of eNOS on different lysates with Rabbit anti-eNOS antibody at 1/5,000 dilution. Lane 1: EA.hy926 cell lysate (20 µg/Lane), Lane 2: HeLa cell lysate (negative) (20 µg/Lane), Lane 3: bEnd.3 cell lysate (20 µg/Lane), Lane 4: RAW264.7 cell lysate (negative) (20 µg/Lane), Lane 5: Mouse placenta tissue lysate (30 µg/Lane), Lane 6: Mouse kidney tissue lysate (30 µg/Lane), Lane 7: Rat placenta tissue lysate (30 µg/Lane), Lane 8: Rat kidney tissue lysate (30 µg/Lane), Exposure time: Lane 1-2: 10 seconds; Lane 3-8: 3 minutes; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/5,000 dilution was used in primary antibody dilution at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
IHC
Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-eNOS antibody at 1/50 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/50 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
IHC
Immunohistochemical analysis of paraffin-embedded mouse spleen tissue with Rabbit anti-eNOS antibody at 1/50 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/50 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
IHC
Immunohistochemical analysis of paraffin-embedded rat spleen tissue with Rabbit anti-eNOS antibody at 1/50 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/50 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ICC/IF
Immunocytochemistry analysis of EA.hy926 (positive) and HeLa (negative) labeling eNOS with Rabbit anti-eNOS antibody at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-eNOS antibody at 1/50 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.
FC
Flow cytometric analysis of HeLa (left, negative) and EA.hy926 (right, positive) cells labeling eNOS. Cells were fixed and permeabilized. Then stained with the primary antibody (1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
IP
eNOS was immunoprecipitated from 0.2 mg bEnd.3 cell lysate with Rabbit anti-eNOS antibody at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate usingRabbit anti-eNOS antibody at 1/5,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: bEnd.3 cell lysate (input), Lane 2: Rabbit anti-eNOS antibody IP in bEnd.3 cell lysate, Lane 3: Rabbit IgG instead of Rabbit anti-eNOS antibody in bEnd.3 cell lysate, Blocking/Dilution buffer: primary antibody dilution, Exposure time: 3 minutes.| Application Notes | WB:1:5000 IHC:1:50-1:200 ICC/IF:1:50 FC:1:1000 IP:1-2μg/sample |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
