Merck, 482728-100UL, Anti-Nitric Oxide Synthase (1131-1144), Inducible Rabbit pAb

General description
Rabbit polyclonal antibody supplied as undiluted serum. Recognizes the ~130 kDa inducible nitric oxide synthase (iNOS) protein.
Recognizes the ~130 kDa iNOS protein.
This Anti-Nitric Oxide Synthase, Inducible Rabbit pAb is validated for use in Frozen Sections, WB, ICC, Paraffin Sections for the detection of Nitric Oxide Synthase, Inducible.
Immunogen
Mouse
a synthetic peptide (CKKGSALEERKATRL) corresponding to amino acids 1131-1144 of mouse macrophage inducible nitric oxide synthase (iNOS), conjugated to KLH
Application
Frozen Sections (1:500)
Immunoblotting (1:2000)
Immunocytochemistry (1:500)
Paraffin Sections (1:500)
Physical form
Undiluted serum.
Preparation Note
Following initial thaw, aliquot and freeze (-20°C).
Analysis Note
Positive Control
Raw 264.7 cells treated with IFNγ and LPS
Other Notes
Exhibits no cross-reactivity with bNOS or eNOS. Variables associated with assay conditions will dictate the proper working dilution.

Recommended Procedure for Immunocytochemistry of Anti-iNOS:
1. Grow cells in appropriate medium.
2. Induce NOS by pretreating cells for 16 h with fresh media containing either: no addition (control), interferon-γ (50 ng/ml), LPS (30 µg/ml) or interferon plus LPS.
3. Wash cells 3-5 min with phosphate buffered saline (PBS).
4. Permeabilize with 2% saponin for 10 min at room temperature.
5. Fix in 4% formalin for 10 min at room temperature.
6. Wash 3-5 min with PBS.
7. Incubate for 20 min at room temperature with anti-iNOS diluted 1:500 in PBS.
8. Wash 3-5 min with PBS.
9. Incubate with biotinylated secondary antibody (i.e. biotin-conjugated goat anti-rabbit IgG, Cat. No. 401313). Wash; incubate with avidin-conjugated alkaline phosphatase (Cat. No. 189726 or 189732).
10. Develop with Naphthol AS Phosphate/Fast Red Violet LB (Cat. No. 477758) or Naphthol AS Phosphate/New Fuchsin (Cat. No. 477759).
Lowenstein, C., et al. 1992. Proc. Natl. Acad. Sci. USA89, 6711.
Lyons, C., et al. 1992. J. Biol. Chem.267, 6370.
Bredt, D.S., et al. 1991. Nature351, 714.
Legal Information
CALBIOCHEM is a registered trademark of Merck KGaA, Darmstadt, Germany
Disclaimer
Toxicity: Standard Handling (A)