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S7150 OxyBlot Protein Oxidation Detection Kit

S7150
200 assays  
Purchase on Sigma-Aldrich

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Tableau de caractéristiques principal

Key Applications
WB
Description
Catalogue NumberS7150
Brand Family Chemicon®
Trade Name
  • OxyBlot
  • Chemicon
DescriptionOxyBlot Protein Oxidation Detection Kit
OverviewOxidative modification of proteins by oxygen free radicals and other reactive species occurs in physiologic and pathologic processes. As a consequence of the modification, carbonyl groups are introduced into protein side chains by a site-specific mechanism. The OxyBlot Kit provides reagents for simple and sensitive immunodetection of these carbonyl groups, which is a hallmark of the oxidation status of proteins.
References
Product Information
Detection methodChemiluminescence
HS Code3002 15 90
Quality LevelMQ100
Applications
ApplicationThe OxyBlot Protein Oxidation Detection Kit provides the reagents to perform the immunoblot detection of carbonyl groups introduced into proteins by oxidative reactions with ozone or oxides of nitrogen or by metal catalyzed oxidation.
Key Applications
  • Western Blotting
Application NotesPreparation of Cell Lysate

1. Wash adherent cells twice in the dish or flask with ice-cold PBS and drain off PBS. Wash non-adherent cells in PBS and centrifuge at 800 to 1000 rpm in a table-top centrifuge for 5 minutes to pellet the cells.
2. Add ice-cold RIPA buffer to cells (1 ml per 107 cells/100 mm dish/150 cm2 flask; 0.5 ml per 5 x 106 cells/60 mm dish/75 cm2 flask).
3. Scrape adherent cells off the dish or flask with either a rubber policeman or a plastic cell scraper that has been cooled in ice-cold distilled water. Transfer the cell suspension into a centrifuge tube. Gently rock the suspension on either a rocker or an orbital shaker in the cold room for 15 minutes to lyse cells.
4. Centrifuge the lysate at 14,000 x g in a precooled centrifuge for 15 minutes. Immediately transfer the supernatant to a fresh centrifuge tube and discard the pellet.
5. Dilute the cell lysate at least 1 : 10 before determining the protein concentration because of the interference of the detergents in the lysis buffer with the Coomassie-based reagent. At this step, the sample can be divided into aliquots and stored at -20¡C for up to a month.
TIP: When working with large volumes of non-adherent cells, the cells may not be cooled quickly enough to maintain the activity of the protein being studied. In this case, pour the cell suspension into a mixture of an equal mass of 2 x PBS and ice, then collect the cells by centrifugation and perform the lysis as described above.

RIPA lysis buffer:
Catalog number 20-188
http://www.millipore.com/catalogue/item/20-188

Reducing agent is very important for preventing oxidation:
The protein lysate may be prepared using a variety of protocols. Most standard buffers such as RIPA and Triton are also suitable. It is recommended that a reducing agent be added to the lysis buffer to prevent the oxidation of proteins that may occur after cell lysis. Lysis buffer containing either 1-2% 2-mercaptoethanol or 50 mM DTT sufficiently inhibits this oxidation, but has no adverse effect on the derivatization reaction in the OxyBlot™ protocol. Purified protein samples are also suitable for analysis with the OxyBlot™ Kit.

For general information about protein extraction, please check this link.

http://www.millipore.com/immunodetection/id3/proteinextraction
Biological Information
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Product Usage Statements
Usage Statement
  • Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Storage and Shipping Information
Storage ConditionsStore the protein standard (component 90450) at -25° to -15°C upon receipt. Store all other components at 2°C to 8°C.
Packaging Information
Material Size200 assays
Transport Information
Supplemental Information
Specifications
Global Trade Item Number
Référence GTIN
S7150 08436037122958

Documentation

OxyBlot Protein Oxidation Detection Kit FDS

Titre

Fiche de données de sécurité des matériaux (FDS) 

Références bibliographiques

Aperçu de la référence bibliographiqueEspèceNº PubMed
Neuroglial alterations in rats submitted to the okadaic acid-induced model of dementia.
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Behavioural brain research  226  420-7. Epub 2011 Oct 1.  2011

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Mononuclear iron enzymes are primary targets of hydrogen peroxide stress.
Adil Anjem,James A Imlay
The Journal of biological chemistry  287  2011

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Autophagy proteins LC3B, ATG5 and ATG12 participate in quality control after mitochondrial damage and influence lifespan.
Sören Mai,Britta Muster,Jürgen Bereiter-Hahn,Marina Jendrach
Autophagy  8  2011

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22170153 22170153
Aerobic exercise training upregulates skeletal muscle calpain and ubiquitin-proteasome systems in healthy mice.
Telma F Cunha,Jose B N Moreira,Nathalie A Paixão,Juliane C Campos,Alex W A Monteiro,Aline V N Bacurau,Carlos R Bueno,Julio C B Ferreira,Patricia C Brum
Journal of applied physiology (Bethesda, Md. : 1985)  112  2011

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Impact of obesity control on circulating level of endothelial progenitor cells and angiogenesis in response to ischemic stimulation.
Yung-Lung Chen,Chia-Lo Chang,Cheuk-Kwan Sun,Chiung-Jen Wu,Tzu-Hsien Tsai,Sheng-Ying Chung,Sarah Chua,Kuo-Ho Yeh,Steve Leu,Jiunn-Jye Sheu,Fan-Yen Lee,Chia-Hung Yen,Hon-Kan Yip
Journal of translational medicine  10  2011

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22568992 22568992
Intra-coronary administration of tacrolimus markedly attenuates infarct size and preserves heart function in porcine myocardial infarction.
Sarah Chua,Steve Leu,Jiunn-Jye Sheu,Yu-Chun Lin,Li-Teh Chang,Ying-Hsien Kao,Chia-Hung Yen,Tzu-Hsien Tsai,Yung-Lung Chen,Hsueh-Wen Chang,Cheuk-Kwan Sun,Hon-Kan Yip
Journal of inflammation (London, England)  9  2011

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Novel protective mechanisms for S-adenosyl-l-methionine against acetaminophen hepatotoxicity: Improvement of key antioxidant enzymatic function.
James Michael Brown,John G Ball,Michael Scott Wright,Stephanie Van Meter,Monica A Valentovic
Toxicology letters  212  2011

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22683606 22683606
Exercise training prevents oxidative stress and ubiquitin-proteasome system overactivity and reverse skeletal muscle atrophy in heart failure.
Telma F Cunha,Aline V N Bacurau,Jose B N Moreira,Nathalie A Paixão,Juliane C Campos,Julio C B Ferreira,Marcelo L Leal,Carlos E Negrão,Anselmo S Moriscot,Ulrik Wisløff,Patricia C Brum
PloS one  7  2011

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22870245 22870245
Chronic paracetamol treatment influences indices of reactive oxygen species accumulation in the aging Fischer 344 X Brown Norway rat aorta.
Kevin M Rice,Sarath Meduru,Sunil K Kakarla,Anjaiah Katta,Sriram P Mupparaju,Brent Kidd,Lynne J Goebel,Eric R Blough
Annals of clinical and laboratory science  42  2011

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22585611 22585611
Enhancement of proteasome function by PA28α overexpression protects against oxidative stress.
Li J, Powell SR, Wang X
The FASEB journal : official publication of the Federation of American Societies for Experimental Biology  25  883-93. Epub 2010 Nov 23.  2010

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21098724 21098724