SP5116P

antibody from Acris Antibodies GmbH

Targeting: NOS1 nNOS, NOS

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

SP5116P

Provider

Acris Antibodies GmbH

Proper citation

Acris Antibodies GmbH Cat#SP5116P, RRID:AB_1001073

Product name

anti NOS1

Antibody type

Polyclonal

Antigen

Synthetic peptide corresponding to residues C N(1411) R L R S E S I A F I E E S K(1425) of human nNOS.

Reactivity

Human, Mouse, Rat

Host

Rabbit

Vial size

0.1 mg

Concentration

1.0 mg/ml

Western blot

Supportive validation

Submitted by

Acris Antibodies GmbH (provider)

Main image

Experimental details

Western blot analysis of nNOS was performed by loading 25 ug of rat brain cell lysates onto an SDS polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked at 4ºC overnight. The membrane was probed with a nNOS polyclonal antibody (Cat.-No SP5116P) at a dilution of 1:200 overnight at 4°C, washed in TBST, and probed with an HRP-conjugated secondary antibody for 1 hr at room temperature in the dark. Chemiluminescent detection was performed using Pierce ECL Plus Western Blotting Substrate. Results show a band at approx. 160 kDa.

Supportive validation

Submitted by

Acris Antibodies GmbH (provider)

Main image

Experimental details

Western blot analysis of nNOS was performed by loading 25 ug of mouse brain cell lysates onto an SDS polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked at 4ºC overnight. The membrane was probed with a nNOS polyclonal antibody (Cat.-No SP5116P) at a dilution of 1:500 overnight at 4°C, washed in TBST, and probed with an HRP-conjugated secondary antibody for 1 hr at room temperature in the dark. Chemiluminescent detection was performed using Pierce ECL Plus Western Blotting Substrate. Results show a band at approx. 160 kDa.

Immunocytochemistry

Supportive validation

Submitted by

Acris Antibodies GmbH (provider)

Main image

Experimental details

Immunofluorescent analysis of nNOS (green) showing staining in the cytoplasm and membrane of U251 cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with an nNOS polyclonal antibody (Cat.-No SP5116P) in 3% BSA-PBS at a dilution of 1:100 and incubated overnight at 4ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-Actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.

Immunohistochemistry

Supportive validation

Submitted by

Acris Antibodies GmbH (provider)

Main image

Experimental details

Immunohistochemistry analysis of nNOS showing positive staining in the cytoplasm of paraffin-treated Human skeletal muscle (right) compared with a negative control in the absence of primary antibody (left). To expose target proteins, antigen retrieval method was performed using 10mM sodium citrate (pH 6.0) microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a nNOS polyclonal antibody (Cat.-No SP5116P) diluted by 3% BSA-PBS at a dilution of 1:200 overnight at 4°C in a humidified chamber. Tissues were washed extensively PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.