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MAB3560

Sigma-Aldrich

Anti-8-Oxoguanine Antibody, clone 483.15

ascites fluid, clone 483.15, Chemicon®

Synonym(s):

8-oxoG

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About This Item

UNSPSC Code:
12352203
eCl@ss:
32160702
NACRES:
NA.41

biological source

mouse

Quality Level

antibody form

ascites fluid

antibody product type

primary antibodies

clone

483.15, monoclonal

species reactivity

monkey, human, primate

species reactivity (predicted by homology)

mouse, rat

manufacturer/tradename

Chemicon®

technique(s)

ChIP: suitable
ELISA: suitable
immunocytochemistry: suitable

isotype

IgM

shipped in

dry ice

target post-translational modification

unmodified

Gene Information

human ... OGG1(4968)

General description

8-Oxoguanine is a mutagenic oxidative damage product of guanine. Oxidatively damaged base 8-oxoguanine is generated in the DNA of all living organisms due to the presence of reactive oxygen species in cells. DNA damage threatens the health of the genetic information stored in every DNA molecule; however enzymes exist in cells to protect against the mutagenic effect of this lesion. 8-oxoguanine is one of the most abundant and well-characterized DNA lesions generated by oxidative stress. It has been estimated that ~180 guanines are oxidized to 8-oxoG per mammalian cell per day. 8-oxoG is a miscoding lesion that can cause G:C to T:A or T:A to G:C transversion mutations. This lesion accumulates in DNA with age and it has been loosly linked to several cancers and diseases, such as Alzheimer′s and Parkinson′s.

Specificity

8-oxoguanine. By competitive ELISA the monoclonal showed no reactivity with dGMP, dAMP, dCMP or TMP in micromolar concentrations. Competition was only observed when the concentrations were increased to the millimolar range.

Immunogen

8-oxoguanine adsorbed on to alumina

Application

Anti-8-Oxoguanine Antibody, clone 483.15 detects level of 8-Oxoguanine & has been published & validated for use in ELISA & IC.
Immunocytochemistry:
On HeLa and Cos7 cells fixed with paraformaldehyde. 8-oxoguanine has been localized to the nucleus in nutrient-deprived cells.

ELISA:
Cell grown in slides were extracted twice for 30 sec with cold 560nM NaCl;0.1% (v/v) Triton X-100;0.02% (w/v) SDS;10mM phosphate buffer pH 7.4 and fixed with freshly prepared 4% PFA for 5-20 minutes at room temperature. {Conlon, KA et al (2000) J. Histotechnology 23(1):37-44}. Typical staining shows that nuclei from extracted cells have define periphery and areas of condensed chromatin. Clone 483.15 staining showed specific but faint nuclear fluorescence staining in cells incubated in supplemented DMEM, but in cells incubated in nutrient-free defined solt solution (NFDSS) {1.8mM calcium chloride, 110mM NaCl, 44mM sodium biocarbonate, pH 7.5} showed strong nuclear fluorescence staining that appeared punctate and gernerally distributed. Fluorescence staining disappears to background levels in cells incubated in nutrient medias even after initial NFDSS treatments {Conlon et al}.

A previous lot of this antibody was used in an ELISA assay.

Optimal working dilutions must be determined by end user.

Quality

Routinely evaluated by immunocytochemistry on NIH/3T3 cells..

Immunocytochemistry:
Confocal fluorescent analysis of NIH/3T3 cells using anti-8-oxoguanine mouse monoclonal antibody.

Physical form

Unpurified mouse monoclonal IgM liquid in buffer containing no preservative.

Storage and Stability

Stable for 6 months at -20ºC in undiluted aliquots from date of receipt.
Handling Recommendations: Upon receipt, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgM and affect product performance.

Analysis Note

Control
Nutrient starved HeLa or Cos7 cells

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Legal Information

CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany

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Storage Class Code

10 - Combustible liquids

WGK

nwg

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable


Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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Due to extensive use in consumer goods, it is important to understand the genotoxicity of silver nanoparticles (AgNPs) and identify susceptible populations. 8-Oxoguanine DNA glycosylase 1 (OGG1) excises 8-oxo-7,8-dihydro-2-deoxyguanine (8-oxoG), a pro-mutagenic lesion induced by oxidative stress. To understand whether
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