- iNOS antibody (ab3523)

IHC with a indirect immunoalkaline phosphatase (APAAP) technique using ab3523. Shows iNOS expression within vessels walls in the Locus Coeruleus after death from septic shock. Picture kindly given to us by Prof. Djillali Annane.

Western blot - iNOS antibody (ab3523)

All lanes : Anti-iNOS antibody (ab3523) at 1/3000 dilution

Lane 1 : Whole tissue lysate prepared from mouse lungs
Lane 2 : Whole tissue lysate prepared from mouse lungs

Lysates/proteins at 20 µg per lane.

Secondary
Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP) (ab6721) at 1/10000 dilution
developed using the ECL technique

Observed band size : 131 kDa (why is the actual band size different from the predicted?)


Exposure time : 30 seconds

This image is courtesy of an anonymous Abreview

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-iNOS antibody (ab3523)

ab3523 staining iNOS in murine lung tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).

Tissue was fixed in paraformaldehyde, blocked with 5% BSA for 30 minutes and then incubated with ab3523 at a 1/50 dilution for 18 hours at 4°C. The secondary used was an undiluted HRP conjugated goat polyclonal.

Image courtesy of an anonymous Abreview.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-iNOS antibody (ab3523)

Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized human eart tissue. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1/200 with a rabbit polyclonal antibody recognizing iNOS (ab3523) or without primary antibody (negative control) overnight at 4ºC in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-iNOS antibody (ab3523)

Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized human lung tissue. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH 6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1/200 with a rabbit polyclonal antibody recognizing iNOS (ab3523) or without primary antibody (negative control) overnight at 4ºC in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.