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Our Abpromise guarantee covers the use of ab15323 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-Fr||Use at an assay dependent concentration. PubMed: 18523253|
|Electron Microscopy||Use at an assay dependent concentration. PubMed: 21071693|
|WB||Use at an assay dependent concentration. Detects a band of approximately 140 kDa (predicted molecular weight: 131 kDa).
Please use at a assay dependent concentration. 1µg/ml was used of a 0.2mg/ml stock (lot GR126616-1). Otherwise 1/250 is recommended for any batches at lower concentration.
|Flow Cyt||Use at an assay dependent concentration.|
|IHC-P||1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|ICC/IF||Use at an assay dependent concentration. PubMed: 20584290|
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.
ab15323 staining macrophages in mouse spleen sections by IHC-P. The tissue was fixed with formaldehyde and a heat mediated antigen retrival step was performed with citric acid pH 6. Blocking of the sample was done with 1%BSA for 10 minutes at 21°C, followed by staining with ab15323 at 1/50 in TBS/BSA/azide for 16h at 21°C. A biotinylated goat anti-rabbit polyclonal antibody at 1/200 was used as the secondary antibody.
ab15323 staining iNOS in Mouse NIH3T3 cell line by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.1% Triton and blocked with 10% serum for 30 minutes at 24°C. Samples were incubated with primary antibody (1/100) for 16 hours at 4°C. An Alexa Fluor® 488-conjugated Donkey anti-rabbit polyclonal was used as the secondary antibody (1/500).
ab15323 staining rat liver (lower image) and spleen (upper image) sections by IHC-P. The tissue was fixed with formaldehyde and a heat mediated antigen retrival step was performed with citric acid pH 6. Blocking of the sample was done with 1%BSA for 10 minutes at 21°C, followed by staining with ab15323 at 1/100 in TBS/BSA/azide for 16h at 21°C. A biotinylated goat anti-rabbit polyclonal antibody at 1/200 was used as the secondary antibody.
Membrane was blocked in 2% milk for 1 hour at RT. Primary anitbody was incubated at 4°C overnight.
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