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482728

Sigma-Aldrich

Anti-Nitric Oxide Synthase (1131-1144), Inducible Rabbit pAb

liquid, Calbiochem®

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Synonym(s):
Anti-iNOS

biological source

rabbit

Quality Level

antibody form

serum

antibody product type

primary antibodies

clone

polyclonal

form

liquid

contains

≤0.1% sodium azide as preservative

species reactivity

mouse, rat, human

manufacturer/tradename

Calbiochem®

storage condition

OK to freeze
avoid repeated freeze/thaw cycles

isotype

IgG

shipped in

wet ice

storage temp.

−20°C

target post-translational modification

unmodified

Gene Information

human ... NOS1(4842)

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)

Warning

Toxicity: Standard Handling (A)

Physical form

Undiluted serum.

Reconstitution

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

Storage Class

10 - Combustible liquids

wgk_germany

WGK 1


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