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World's First kit of its kind for research into aging, oxidative stress, and oxidative stress related disease.

Protein Carbonyls Immunohistochemical Staining Kit: for the specific detection of protein oxidation by ROS.


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Background

Reactive oxygen species (ROS) are produced as a result of normal cell metabolism or by exposure to ionizing radiation, chemicals, or other environmental stress. ROS are well known to promote non-specific protein oxidation, with negative effects protein structure and function. Typical ROS-induced protein modifications include the transformation of lysine, arginine, proline, and threonine side chain amines into aminoacyl carbonyls. The chemical stability of these carbonyl derivatives allows their detection and quantification, providing a sensitive and reliable marker of ROS-mediated protein oxidation. In the central feature of this kit, carbonyl groups in tissue are first derivitized by reaction with 2,4-Dinitrophenylhydrazine (DNPH). DNPH-derivitized carbonyls are then detected using an anti-DNP specific antibody suitable for immunohistochemical procedures. The Protein Carbonyls Immunohistochemical Staining Kit is the first kit to enable detection of protein carbonyls by immunohistochemical staining. A detection kit for protein carbonyls in cell lysates by Western blotting with anti-DNP antibody is also available (Cat No.SML-ROIK03-EX).


 Oxidative Stress and Aging Research Kits Flyer [PDF]

Kit component

Tested Application Immunohistochemistry with methacarn-fixed paraffin-embedded tissue (see note 1 below)
Antibody Rabbit anti-DNP for immunohistochemistry
Control Specimen Mouse Kidney, 2 slides, (Methacarn-fixed paraffin-embedded sections)
DNPH solution 2,4-Dinitrophenylhydrazine (DNPH) solution
Storage and Stability Antibody, Specimen, DNPH solution: 1 year at 4°C
Note 1 This kit cannot be used for tissues fixed with aldehyde containg fixatives such as formaldehyde, paraformaldehyde or glutaraldehyde. The use of Methacarn fixative is strongly advised. Methacarn fixing solution: (Methanol/Chloroform/Acetic Acid (6:3:1).
Note 2 This kit does not contain Sodium Azide (NaN3)
Reagents not included Detection reagents for rabbit Ig. (e.g., Elite ABC Kit (Rabbit IgG), Vector Labs)

Mouse liver and kidney sections stained with rabbit anti-DNP antibody at 1:100 dilution and developed by 3,3’-diaminobenzidine (DAB). Specimens were incubated with (DNPH(+)) or without (DNPH(-)) DNPH solution. Protein carbonyls are detected only in DNPH-treated specimen and not detected at all in untreated specimen.
Courtesy of Dr. Akihito Ishigami, Tokyo Metropolitan Institute of Gerontology

<Reference>
1. Nakamura A. et al., Analysis of protein carbonyls with 2,4-dinitrophenyl hydrazine and its antibodies by immunoblot in two-dimensional gel electrophoresis. J Biochem (Tokyo)119 768-774 (1996)
2. Goto S. et al., Age-associated, oxidatively modified proteins: A critical evaluation. Age 20 81-89 (1997)
3. Goto S. et al., Carbonylated Proteins in Aging and Exercise: Immunoblot Approaches. Mech Ageing Dev 107 245-253 (1999)
4. Nakamura A. et al., Vitellogenin-6 is a major carbonylated protein in aged nematode, Caenorhabditis elegansBiochem Biophys Res Commun. 264 580-583 (1999)
5. Robinson CE. et al., Determination of protein carbonyl groups by immunoblotting. Anal Biochem. 266 48-57 (1999)
6. Sato T. et al., Senescence marker protein-30 protects mice lungs from oxidative stress, aging, and smoking.Am J Respir Crit Care Med174 530-537 (2006)

Product List

Product Name Cat# Quantity Price

Protein Carbonyls Immunohistochemical Staining Kit (50 slides) DataSheet ,

SML-ROIK04-EX

1 KIT ¥50,000
$667

Protein carbonyls western blot detection kit(15Blots) (Rabbit)  DataSheet

SML-ROIK03-EX

1 KIT see also
# SML-ROIK032EX
Size 1

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