Anti-iNOS antibody [EPR16635] - BSA and Azide free

• WB

Lab

Western blot - Anti-iNOS antibody [EPR16635] - BSA and Azide free (AB213987)

This data was developed using the same antibody clone in a different buffer formulation (ab178945). Blocking and diluting buffer 5% NFDM/TBST. iNOS is not normally expressed in the brain, but can be induced in the brain after inflammatory, infectious, or other damages (PMID : 11138926, PMID : 16156895, PMID : 10322315).

All lanes:

Western blot - Anti-iNOS antibody [EPR16635] (<a href='/en-us/products/primary-antibodies/inos-antibody-epr16635-ab178945'>ab178945</a>) at 1/1000 dilution

Lane 1:

Mouse hippocampus tissue lysate at 20 µg

Lane 2:

Mouse colon tissue lysate at 20 µg

Lane 3:

Mouse colon cancer tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 131 kDa

false

Exposure time: 180s

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-iNOS antibody [EPR16635] - BSA and Azide free (AB213987)

Immunocytochemistry/Immunofluorescence analysis of RAW 264.7 non-treated and LPS treated (0.1 μg/mL) cells labelling iNOS with ab178945 at 1/250. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. The cells were co-stained with ab195889, Alexa Fluor^® 594 conjugated anti-alpha tubulin (1/200). Nuclei counterstained with DAPI (blue).

Secondary antibody only controls performed on non-treated and treated cells.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab178945).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-iNOS antibody [EPR16635] - BSA and Azide free (AB213987)

Clone EPR16635 (ab213987) has been successfully conjugated by Abcam. This image was generated using Anti-iNOS antibody [EPR16635] (Alexa Fluor® 647). Please refer to ab209027 for protocol details.

ab209027 staining iNOS in Raw264.7 cells. The lower panel shows cells treated with 1ug/ml Lipopolysaccharides (24 hr).  The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab209027at 1/100 dilution (shown in red) and ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor^® 488), at 1/250 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

This product also gave a positive signal under the same testing conditions in SW480 cells fixed with 4% formaldehyde (10 min).

• I-ELISA

Supplier Data

Indirect ELISA - Anti-iNOS antibody [EPR16635] - BSA and Azide free (AB213987)

ELISA analysis of House mouse iNOS recombinant protein at 1000 ng/mL with ab178945. An Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1/2500 dilution was used as the secondary antibody.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab178945).

• IP

Supplier Data

Immunoprecipitation - Anti-iNOS antibody [EPR16635] - BSA and Azide free (AB213987)

iNOS was immunoprecipitated from 1mg of RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate treated with 1 μg/ml LPS for 24h with ab178945 at 1/100 dilution.

Lane 1 : RAW 264.7 whole cell lysate treated with 1 μg/ml LPS for 24h,10ug (Input).

Lane 2 : ab178945 IP in RAW 264.7 whole cell lysate treated with 1 μg/ml LPS for 24h.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab178945 in RAW 264.7 whole cell lysate treated with 1 μg/ml LPS for 24h.

Western blot was performed from the immunoprecipitate using ab178945 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1500 dilution

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 30 seconds.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab178945).

All lanes:

Immunoprecipitation - Anti-iNOS antibody [EPR16635] (<a href='/en-us/products/primary-antibodies/inos-antibody-epr16635-ab178945'>ab178945</a>)

Predicted band size: 131 kDa

false

• WB

Supplier Data

Western blot - Anti-iNOS antibody [EPR16635] - BSA and Azide free (AB213987)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab178945).

All lanes:

Western blot - Anti-iNOS antibody [EPR16635] - BSA and Azide free (ab213987) at 1/500 dilution

Lane 1:

HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysates at 15 µg

Lane 2:

HepG2 (Human hepatocellular carcinoma epithelial cell) treated with 10ug/ml lipopolysaccharides for 6 hours whole cell lysates at 15 µg

Lane 3:

THP-1 (Human monocytic leukemia monocyte) whole cell lysates at 15 µg

Lane 4:

THP-1 (Human monocytic leukemia monocyte) treated with 100ng/ml lipopolysaccharides for 3 hours whole cell lysates at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/200000 dilution

Predicted band size: 131 kDa

false

Exposure time: 3min

• WB

Lab

Western blot - Anti-iNOS antibody [EPR16635] - BSA and Azide free (AB213987)

Blocking and dilution buffer : 5% NFDM/TBST.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab178945).

All lanes:

Western blot - Anti-iNOS antibody [EPR16635] - BSA and Azide free (ab213987) at 1/20000 dilution

Lane 1:

Untreated RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate at 20 µg

Lane 2:

RAW 264.7 whole cell lysate treated with 0.1 µg/mL LPS for 6 hours at 20 µg

Secondary

All lanes:

Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 131 kDa

Observed band size: 131 kDa

false

Exposure time: 30s