eNOS Recombinant Rabbit Monoclonal Antibody [PSH16-96] - BSA and Azide free

☑ Relative expression (RE)

This data was developed using HA723892, the same antibody clone in a different buffer formulation.
Western blot analysis of eNOS on different lysates with Rabbit anti-eNOS antibody (HA723892) 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)

Predicted band size: 133 kDa
Observed band size: 133 kDa

Exposure time: Lane 1-2: 10 seconds; Lane 3-8: 3 minutes; ECL: K1801;

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 (HA723892) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

☑ Relative expression (RE)

This data was developed using HA723892, the same antibody clone in a different buffer formulation.
Immunocytochemistry analysis of EA.hy926 (positive) and HeLa (negative) labeling eNOS with Rabbit anti-eNOS antibody (HA723892) 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 (HA723892) at 1/50 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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 (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

This data was developed using HA723892, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Rabbit anti-eNOS antibody (HA723892) at 1/200 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 (HA723892) at 1/200 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.

This data was developed using HA723892, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded human placenta tissue with Rabbit anti-eNOS antibody (HA723892) at 1/200 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 (HA723892) at 1/200 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.

This data was developed using HA723892, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-eNOS antibody (HA723892) 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 (HA723892) 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.

This data was developed using HA723892, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-eNOS antibody (HA723892) 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 (HA723892) 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.

This data was developed using HA723892, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded mouse spleen tissue with Rabbit anti-eNOS antibody (HA723892) 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 (HA723892) 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.

This data was developed using HA723892, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-eNOS antibody (HA723892) 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 (HA723892) 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.

This data was developed using HA723892, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded rat spleen tissue with Rabbit anti-eNOS antibody (HA723892) 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 (HA723892) 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.

This data was developed using HA723892, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded rat placenta tissue with Rabbit anti-eNOS antibody (HA723892) at 1/200 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 (HA723892) at 1/200 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.

☑ Relative expression (RE)

This data was developed using HA723892, the same antibody clone in a different buffer formulation.
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 (HA723892, 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 iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

This data was developed using HA723892, the same antibody clone in a different buffer formulation.
Western blot analysis of eNOS on different lysates with Rabbit anti-eNOS antibody (HA723892) at 1/5,000 dilution.

Lane 1: His-tagged eNOS recombinant protein
Lane 2: His-tagged nNOS recombinant protein

Lysates/proteins at 20 ng/Lane.

Exposure time: 2 seconds; ECL: K1801;

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 (HA723892) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

This data was developed using HA723892, the same antibody clone in a different buffer formulation.
eNOS was immunoprecipitated from 0.2 mg bEnd.3 cell lysate with HA723892 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA723892 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: HA723892 IP in bEnd.3 cell lysate
Lane 3: Rabbit IgG instead of HA723892 in bEnd.3 cell lysate

Blocking/Dilution buffer: primary antibody dilution (K1803)
Exposure time: 3 minutes; ECL: K1801