		Technical Data
C9002-75	Cysteine, S-Nitrosylated (SNO-Cys)

Description:
Nitric oxide (NO), generated by cell type-specific NO-synthase (NOS) isoforms, is a freely diffusible intercellular messenger that functions in target cells in NOS-dependent signaling. S-nitrosylation of cysteine thiols in proteins by the highly labile NO radical has been identified as an important effector of NO-related bioactivity both in NOS-containing cells and intercellular signaling. Most cells contain low levels of nitrosylated proteins that are thought to be regulated by S-nitrosylation and denitrosylation. S-nitrosylation of proteins serves as a ubiquitous post-translational modification that dynamically regulates a broad functional spectrum of proteins. The majority of these proteins are regulated by S-nitrosylation on a single critical cysteine residue within an acidic/basic or hydrophobic structural motif that may also be subject to oxygen- or glutathione-dependent modification. NO-sensitive ion channels including the cardiac and skeletal muscle ryanodine receptor (RyR1), N-methyl-D-aspartate receptor (NMDAR) complex, cyclic-nucleotide gated ion channel, are modulated by S-nitrosylation. S-nitrosylation of capsase-3 inhibits apoptosis signaling. S-nitrosylation activates matrix metalloproteinase-9 (MMP-9) and induces neuronal apoptosis. The small G-protein p21Ras and Jun kinase are regulated by S-nitrosylation. The activity of transcription factors such as NFkB, c-jun, and c-fos is modulated by S-nitrosylation. In addition, the formation of S-nitrosylated glutathione (GSNO) has been proposed to be one of the major storage forms of NO in vivo.

Applications:
Suitable for use in Immunohistochemistry, Western Blot and ELISA. Other applications not tested.

Recommended Dilutions:
Immunohistochemistry: 2-10ug/ml. Use bovine endothethial cells treated with Ca+2 ionophore A23187 as a control.
Western Blot: 1:500-1:2000 using nitrosylated-Cysteine–BSA (C9002-75A) as a control.
ELISA: 1:1000-1:5000; using 50-100ng control proteins/well.
Optimal dilution to be determined by researcher.

Storage and Stability:
Lyophilized powder may be stored at -20°C. Stable for 12 months at -20°C. Reconstitute with sterile PBS. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Reconstituted product is stable for 12 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

Type	Isotype	Clone	Grade
Pab	IgG		Affinity Purified

Size		Storage	Shipping	Source	Host
100ug		-20°C	Blue Ice		Rabbit

Concentration:

~1mg/ml (after reconstitution)

Immunogen:

Nitrosylated-Cystein, conjugated to KLH (hemocyanin).

Purity:

Purified by immunoaffinity chromatography.

Form

Supplied as a lyophilized powder. Reconstitute with 100ul sterile PBS.

Specificity:

Recognizes nitrosylated-cysteine modified proteins from all species. No significant reactivity is observed with unmodified BSA or Cysteine-BSA (not nitrosylated).

Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.

1. Sunico, C.R., J. Neurosci. 25: 1448-1458 (2005). 2. Gow, A.J., et al., JBC 158: 308-314 (2002). 3. Sun, J., et al., PNAS 98: 11,158-11,162 (2001). 4. Gu, Z., et al., Science 297: 1186-1190 (2002). 5. Park, H.S., et al., PNAS 97: 14,382-14,387 (2000).