Product Specification:
| Alternative Name: | iNOS, NOS II |
| Host: | Rabbit |
| Immunogen: | Synthetic peptide corresponding to aa 1131-1144 (CK1131KGSALEEPKATRL1144) of mouse inducible nitric oxide synthase (iNOS; NOS II). |
| UniProt ID: | P29477 |
| Gene/Protein Identifier: | NM_010927 (RefSeq) |
| Species reactivity: | Mouse |
| Crossreactivity: | Does not cross-react with eNOS (NOS III) and nNOS (NOS I). |
| Applications: | IF, IHC (PS), WB |
| Recommended Dilutions/Conditions: | Immunofluorescence (1:100-1:1,000) Immunohistochemistry (paraffin sections, 1:200) Immunohistochemistry (frozen sections, 1:500) Western Blot (1:2,000) Suggested dilutions/conditions may not be available for all applications. Optimal conditions must be determined individually for each application. |
| Application Notes: | Detects a band of ~130kDa by Western blot. |
| Formulation: | Liquid. Antiserum diluted in PBS containing 0.05% sodium azide. |
| Handling: | Avoid freeze/thaw cycles. |
| Shipping: | Shipped on Blue Ice |
| Long Term Storage: | -20°C |
| Protocol: | Western blot of NOS 1. For the study of iNOS, prepare cytosolic extracts from cells that are not treated and do not express iNOS activity (control) or cells induced to synthesize NO by pretreatment for 16 hours with a combination pf LPS (30μg/ml) and interferon-γ (50ng/ml). 2. Run cell extracts on SDS-PAGE using a 4-12% TRIS-glycine gel. Transfer to blotting membrane. 3. Block membrane in 5%dried milk in PBS containing 1% Tween-20 (PBS-T). 4. Wash 3 x 5 minutes in PBS. 5. Incubate for 1 hour at room temperature with antibody to NOS diluted in PBS. 6. Wash 3 x 5 minutes in PBS. 7. Incubate for 1 hour with secondary antibody to rabbit coupled to biotin, then with streptavidin conjugated to alkaline phosphatase. 8. Wash 3 x 5 minutes in PBS. 9. React to effect with BCIP or Lumi-Phos 530
Immunohistochemical Detection of iNOS using PAb to iNOS |
