Anti-iNOS 抗体

• WB

Lab

Western blot - Anti-iNOS antibody (AB15323)

This blot was produced using a 4-12% Bis-Tris gel under the MOPS buffer system under denaturing, reducing conditions. The gel was run at 200V for 60 minutes before being transferred onto a nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour before being incubated with rabbit polyclonal to iNOS (ab15323; 1 ug x mL-1 (based on antibody concentration of 0.2mg/ml - lot GR126616-1) and the loading control mouse anti-GAPDH antibody (ab8245; 1 : 10000) overnight at 4°C. Antibody binding was detected using infrared (IR) labelled goat anti-rabbit (green; 1 : 10000) and IR-goat anti-mouse (red; 1 : 10000) for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-iNOS antibody (ab15323) at 1 µg/mL

Lane 1:

Raw264.7 at 20 µg

Lane 2:

Raw264.7 + LPS (1 ug per mL; 18 hours) at 20 µg

Secondary

All lanes:

IR- goat anti-rabbit (green) at 1/10000 dilution

Predicted band size: 131 kDa

false

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-iNOS antibody (AB15323)

IHC image of iNOS staining in a section of formalin-fixed paraffin-embedded normal rat lung performed on a Leica BOND^TM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab15323, 1/100 dilution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-iNOS antibody (AB15323)

IHC image of iNOS staining in a section of formalin-fixed paraffin-embedded normal mouse lung performed on a Leica BOND^TM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab15323, 1/100 dilution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-iNOS antibody (AB15323)

Immunohistochemical analysis of formalin fixed paraffin embedded mouse lung labelling iNOS with ab15323 at a concentration of 1µg/ml.

The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a ChromoMap DAB kit and anti-rabbit HQ and anti-HQ HRP detection.
Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32mins.
ab15323 Anti-iNOS antibody was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.
Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual).

• WB

Lab

Western blot - Anti-iNOS antibody (AB15323)

All lanes:

Western blot - Anti-iNOS antibody (ab15323) at 5 µg/mL

Lane 1:

Raw 264.7 whole cell lysate + PMA at 20 µg

Lane 2:

Raw 264.7 whole cell lysate + PMA + LPS + Brefeldin A at 20 µg

Predicted band size: 131 kDa

Observed band size: 140 kDa

true

Exposure time: 8min

• WB

Lab

Western blot - Anti-iNOS antibody (AB15323)

Membrane was blocked in 2% milk for 1 hour at RT. Primary anitbody was incubated at 4°C overnight.

All lanes:

Western blot - Anti-iNOS antibody (ab15323) at 5 µg/mL

Lane 1:

Human recombinant iNOS protein at 0.1 µg

Lane 2:

Mouse recombinant iNOS protein at 0.1 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/50000 dilution

Predicted band size: 131 kDa,47 kDa,68 kDa

true

Exposure time: 1min