MOLECULAR SIGNATURE® Anti-DNA/RNA Damage Antibody [15A3]

Mouse Anti- DNA/RNA Damage Monoclonal IgG2b

Catalog No. SMC-155

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Species Reactivity ALL
Applications WB IHC ICC/IF FCM IP
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SKU: SMC-155 Categories: , .

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SMC-155_DNA-Damage_Antibody_15A3_IHC_Mouse_Retinal-Injury-Model_1.png
Mouse Anti-DNA Damage Antibody [15A3] used in Immunohistochemistry (IHC) on Mouse inflamed colon (SMC-155)Mouse Anti-DNA Damage Antibody [15A3] used in Immunohistochemistry (IHC) on Mouse backskin (SMC-155)Mouse Anti-DNA Damage Antibody [15A3] used in Immunohistochemistry (IHC) on Rat Ischemic fresh brain tissue (SMC-155)
Product Name DNA/RNA Damage Antibody
Description

Mouse Anti- DNA/RNA Damage Monoclonal IgG2b

Species Reactivity Species Independent
Applications IHC, ICC/IF, ELISA, DB
Antibody Dilution IHC (1:1000); optimal dilutions for assays should be determined by the user.
Host Species Mouse
Immunogen 8-hydroxy-guanosine-BSA and –casein conjugates
Concentration 1 mg/ml
Conjugates Alkaline Phosphatase, APC, ATTO 390, ATTO 488, ATTO 565, ATTO 594, ATTO 633, ATTO 655, ATTO 680, ATTO 700, Biotin, FITC, HRP, PE/ATTO 594, PerCP, RPE, Streptavidin, Unconjugated
 PerCP 
Overview:

  • Peridinin-Chlorophyll-Protein Complex
  • Small phycobiliprotein
  • Isolated from red algae
  • Large stokes shift (195 nm)
  • Molecular Weight: 35 kDa

PerCP Datasheet

 PerCP Fluorophore Absorption and Emission Spectrum Optical Properties:

λex = 482 nm

λem = 677 nm

εmax = 1.96 x 106

Laser = 488 nm

 

 PE/ATTO 594
PE/ATTO 594 is a tandem conjugate, where PE is excited at 535 nm and transfers energy to ATTO 594 via FRET (fluorescence resonance energy transfer), which emits at 627 nm.
Overview:

  • High fluorescence yield
  • High photostability
  • Very hydrophilic
  • Excellent solubility in water
  • Very little aggregation

PE/ATTO 594 Datasheet

PE-ATTO 594 Fluorophore Conjugate Excitation and Emission Spectra Optical Properties:

λex = 535 nm

λem = 627 nm

Laser = 488 to 561 nm

 

  FITC (Fluorescein)
Overview:

  • Excellent fluorescence quantum yield
  • High rate of photobleaching
  • Good solubility in water
  • Broad emission spectrum
  • pH dependent spectra
  • Molecular formula: C20H12O5
  • Molar mass: 332.3 g/mol

FITC-Fluorescent-conjugate

FITC Fluorescein Fluorophore Excitation and Emission Spectra Optical Properties:

λex = 494 nm

λem = 520 nm

εmax = 7.3×104

Φf = 0.92

τfl = 5.0 ns

Brightness = 67.2

Laser = 488 nm

Filter set = FITC

 

 ATTO 700
Overview:

  • High fluorescence yield
  • Excellent thermal and photostability
  • Quenched by electron donors
  • Very hydrophilic
  • Good solubility in polar solvents
  • Zwitterionic dye
  • Molar Mass: 575 g/mol

ATTO 700 Datasheet

 ATTO 700 Fluorophore Absorption and Emission Spectrum Optical Properties:

λex = 700 nm

λem = 719 nm

εmax = 1.25×105

Φf = 0.25

τfl = 1.6 ns

Brightness = 31.3

Laser = 676 nm

Filter set = Cy®5.5

 

 ATTO 680
Overview:

  • High fluorescence yield
  • Excellent thermal and photostability
  • Quenched by electron donors
  • Very hydrophilic
  • Good solubility in polar solvents
  • Zwitterionic dye
  • Molar Mass: 631 g/mol

ATTO 680 Datasheet

 ATTO 680 Fluorophore Absorption and Emission Spectrum Optical Properties:

λex = 680 nm

λem = 700 nm

εmax = 1.25×105

Φf = 0.30

τfl = 1.7 ns

Brightness = 37.5

Laser = 633 – 676 nm

Filter set = Cy®5.5

 

 ATTO 655
Overview:

  • High fluorescence yield
  • High thermal and photostability
  • Excellent ozone resistance
  • Quenched by electron donors
  • Very hydrophilic
  • Good solubility in polar solvents
  • Zwitterionic dye
  • Molar Mass: 634 g/mol

ATTO 655 Datasheet

ATTO 655 Fluorophore Absorption and Emission Spectrum Optical Properties:

λex = 663 nm

λem = 684 nm

εmax = 1.25×105

Φf = 0.30

τfl = 1.8 ns

Brightness = 37.5

Laser = 633 – 647 nm

Filter set = Cy®5

 

 ATTO 633
Overview:

  • High fluorescence yield
  • High thermal and photostability
  • Moderately hydrophilic
  • Good solubility in polar solvents
  • Stable at pH 4 – 11
  • Cationic dye, perchlorate salt
  • Molar Mass: 652.2 g/mol

ATTO 633 Datasheet

ATTO 633 Fluorophore Absorption and Emission Spectrum Optical Properties:

λex = 629 nm

λem = 657 nm

εmax = 1.3×105

Φf = 0.64

τfl = 3.2 ns

Brightness = 83.2

Laser = 633 nm

Filter set = Cy®5

 

 ATTO 594
Overview:

  • High fluorescence yield
  • High photostability
  • Very hydrophilic
  • Excellent solubility in water
  • Very little aggregation
  • New dye with net charge of -1
  • Molar Mass: 1137 g/mol

ATTO 594 Datasheet

 ATTO 594 Fluorophore Excitation and Emission Spectrum Optical Properties:

λex = 601 nm

λem = 627 nm

εmax = 1.2×105

Φf = 0.85

τfl = 3.5 ns

Brightness = 102

Laser = 594 nm

Filter set = Texas Red®

 

 ATTO 565
Overview:

  • High fluorescence yield
  • High thermal and photostability
  • Good solubility in polar solvents
  • Excellent solubility in water
  • Very little aggregation
  • Rhodamine dye derivative
  • Molar Mass: 611 g/mol

ATTO 565 Datasheet

 ATTO 565 Fluorophore Excitation and Emission Spectra Optical Properties:

λex = 563 nm

λem = 592 nm

εmax = 1.2×105

Φf = 0.9

τfl = 3.4 n

Brightness = 10

Laser = 532 nm

Filter set = TRITC

 

  ATTO 488
Overview:

  • High fluorescence yield
  • High photostability
  • Very hydrophilic
  • Excellent solubility in water
  • Very little aggregation
  • New dye with net charge of -1
  • Molar Mass: 804 g/mol 

ATTO 488 Datasheet

  ATTO 488 Fluorophore Excitation and Emission Spectra Optical Properties:

λex = 501 nm

λem = 523 nm

εmax = 9.0×104

Φf = 0.80

τfl = 4.1 ns

Brightness = 72

Laser = 488 nm

Filter set = FITC

 

  ATTO 390
Overview:

  • High fluorescence yield
  • Large Stokes-shift (89 nm)
  • Good photostability
  • Moderately hydrophilic
  • Good solubility in polar solvents
  • Coumarin derivate, uncharged
  • Low molar mass: 343.42 g/mol 

ATTO 390 Datasheet

ATTO 390 Fluorescent Dye Excitation and Emission Spectra Optical Properties:

λex = 390 nm

λem = 479 nm

εmax = 2.4×104

Φf = 0.90

τfl = 5.0 ns

Brightness = 21.6

Laser = 365 or 405 nm

 

APC (Allophycocyanin)
Overview:

  • High quantum yield
  • Large phycobiliprotein
  • 6 chromophores per molecule
  • Isolated from red algae
  • Molecular Weight: 105 kDa

APC Datasheet

 APC Fluorophore Absorption and Emission Spectrum Optical Properties:

λex = 650 nm

λem = 660 nm

εmax = 7.0×105

Φf = 0.68

Brightness = 476

Laser = 594 or 633 nm

Filter set = Cy®5

 

Streptavidin

Properties:

  • Homo-tetrameric protein purified from Streptomyces avidinii which binds four biotin molecules with extremely high affinity
  • Molecular weight: 53 kDa
  • Formula: C10H16N2O3S
  • Applications: Western blot, immunohistochemistry, and ELISA

Streptavidin Datasheet

BiotinBiotin Conjugate Structure

Properties:

  • Binds tetrameric avidin proteins including Streptavidin and neuravidin with very high affinity
  • Molar mass: 244.31 g/mol
  • Formula: C10H16N2O3S
  • Applications: Western blot, immunohistochemistry, and ELISA

Biotin Datasheet

HRP (Horseradish peroxidase)

Properties:

  • Enzymatic activity is used to amplify weak signals and increase visibility of a target
  • Readily combines with hydrogen peroxide (H2O2) to form HRP-H2O2 complex which can oxidize various hydrogen donors
  • Catalyzes the conversion of:
    • Chromogenic substrates (e.g. TMB, DAB, ABTS) into coloured products
    • Chemiluminescent substrates (e.g. luminol and isoluminol) into light emitting products via enhanced chemiluminescence (ECL)
    • Fluorogenic substrates (e.g. tyramine, homovanillic acid, and 4-hydroxyphenyl acetic acid) into fluorescent products
  • High turnover rate enables rapid generation of a strong signal
  • 44 kDa glycoprotein
  • Extinction coefficient: 100 (403 nm)
  • Applications: Western blot, immunohistochemistry, and ELISA

HRP Datasheet

AP (Alkaline Phosphatase)

Properties:

  • Broad enzymatic activity for phosphate esters of alcohols, amines, pyrophosphate, and phenols
  • Commonly used to dephosphorylate the 5’-termini of DNA and RNA to prevent self-ligation
  • Catalyzes the conversion of:
    • Chromogenic substrates (e.g. pNPP, naphthol AS-TR phosphate, BCIP) into coloured products
    • Fluorogenic substrates (e.g. 4-methylumbelliferyl phosphate) into fluorescent products
  • Molecular weight: 140 kDa
  • Applications: Western blot, immunohistochemistry, and ELISA

AP Datasheet

  R-PE (R-Phycoerythrin)
Overview:

  • Broad excitation spectrum
  • High quantum yield
  • Photostable
  • Member of the phycobiliprotein family
  • Isolated from red algae
  • Excellent solubility in water
  • Molecular Weight: 250 kDa

R-PE Datasheet

 R-PE Fluorophore Excitation and Emission Spectra Optical Properties:

λex = 565 nm

λem = 575 nm

εmax = 2.0×106

Φf = 0.84

Brightness = 1.68 x 103

Laser = 488 to 561 nm

Filter set = TRITC

 

Properties

Storage Buffer PBS, 50% glycerol, 0.09% sodium azide
Storage Temperature -20ºC
Shipping Temperature Blue Ice or 4ºC
Purification Protein G Purified
Clonality Monoclonal
Clone Number 15A3
Isotype IgG2b
Specificity Recognizes markers of oxidative damage to DNA (8-hydroxy-2’-deoxyguanosine, 8-hydroxyguanine and 8-hydroxyguanosine).
Cite This Product Mouse Anti- DNA Damage Monoclonal, Clone 15A3 (StressMarq Biosciences Inc., Victoria BC CANADA, Catalog # SMC-155)

Biological Description

Alternative Names 8-Hydroxy Guanine Antibody, 8-OH-dG Antibody, 8OHG Antibody, 80G Antibody, 8 hydroxyguanine Antibody, 8 hydroxy 2’ deoxyguanosine Antibody, 8 hydroxyguanosine Antibody, 8 OHG Antibody, 8-OHG Antibody, 8OHdG Antibody
Research Areas Cancer, Cell Signaling, Oxidation, Oxidative Stress, Post-translational Modifications
Scientific Background DNA or RNA damage is due to environmental factors and normal metabolic processes inside the cell, that then hinder the ability of the cell to carry out its functions. There are four main types of DNA due to endogenous cellular processes and they are oxidation, alkylation, hydrolysis and mismatch of the bases. During the oxidation of bases, highly reactive chemical entities collectively known as RONS, occurs. RONS stands for reactive oxygen and nitrogen species and includes nitric oxide, superoxide, hydroxyl radical, hydrogen peroxide and peroxynitrite. Numerous studies have shown that RONS causes a variety of issues including DNA damage(1). 8-hydroxyguanine, 8-hydroxy-2’-deoxyguanonsine and 8- hydroxyguanosine are all RNA and DNA markers of oxidative damage. 8-hydroxy-2’-guanosine is produced by reactive oxygen and nitrogen species including hydroxyl radical and peroxynitrite. Specifically its high biological relevance is due to its ability to induce G to T transversions, which is one of the most frequent somatic mutations (2). 8-hydroxy-guanine has been the most frequently studied type of DNA base damage, with studies in diabetes, and cancer. Base modifications of this type arise from radical-induced hydroxylation and cleavage reactions of the purine ring (3, 4). And finally, 8-hydroxy-guanosine, like 8-hydroxy-2’-guanosine, induces a mutagenic transversion of G to T in DNA. Its role has specifically been tested in the development of diabetes, hypertension and strokes (5, 6, and 7).
References 1. Kim H.W., Murakami A., Williams M.V., and Ohigashi H. (2003) Carcinogenesis 24(2): 235-241.
2. Pilger A. and Rudiger H.W. (2006) Int Arch Occup Environ Health. 80(1): 1-15.
3. Malins D.C. and Haimanot R. (1991) Cancer Res. 51(19): 5430-5432.
4. Kvam E. and Tyrrell R.M. (1997) Carcinogenesis 18(11): 2281-2283.
5. Kowluru R.A., Atasi L., and Ho Y.S. (2006) Invest Ophthalmol Vis Sci 47(4): 1594-9.
6. Bowers R. et al. (2004) Am J Respir Crit Care Med. 169(6): 764-9.
7. Cui J., Holmes E.H., Greene T.G., and Liu P.K. (2000) Faseb J. 14(7): 955-67.

Product Images

<p>Immunohistochemistry analysis using Mouse Anti-DNA Damage Monoclonal Antibody, Clone 15A3 (SMC-155). Tissue: Retinal Injury Model. Species: Mouse. Primary Antibody: Mouse Anti-DNA Damage Monoclonal Antibody (SMC-155) at 1:1000. Secondary Antibody: Alexa Fluor 594 Goat Anti-Mouse (red). Courtesy of: Dr. Rajashekhar Gangaraju, University of Indiana, Department of Opthamology, Eugene and Marilyn Glick Eye Institute.</p>

Immunohistochemistry analysis using Mouse Anti-DNA Damage Monoclonal Antibody, Clone 15A3 (SMC-155). Tissue: Retinal Injury Model. Species: Mouse. Primary Antibody: Mouse Anti-DNA Damage Monoclonal Antibody (SMC-155) at 1:1000. Secondary Antibody: Alexa Fluor 594 Goat Anti-Mouse (red). Courtesy of: Dr. Rajashekhar Gangaraju, University of Indiana, Department of Opthamology, Eugene and Marilyn Glick Eye Institute.

<p>Immunohistochemistry analysis using Mouse Anti-DNA Damage Monoclonal Antibody, Clone 15A3 (SMC-155). Tissue: inflamed colon. Species: Mouse. Fixation: Formalin. Primary Antibody: Mouse Anti-DNA Damage Monoclonal Antibody (SMC-155) at 1:1000000 for 12 hours at 4°C. Secondary Antibody: Biotin Goat Anti-Mouse at 1:2000 for 1 hour at RT. Counterstain: Mayer Hematoxylin (purple/blue) nuclear stain at 200 µl for 2 minutes at RT. Magnification: 40x.</p>

Immunohistochemistry analysis using Mouse Anti-DNA Damage Monoclonal Antibody, Clone 15A3 (SMC-155). Tissue: inflamed colon. Species: Mouse. Fixation: Formalin. Primary Antibody: Mouse Anti-DNA Damage Monoclonal Antibody (SMC-155) at 1:1000000 for 12 hours at 4°C. Secondary Antibody: Biotin Goat Anti-Mouse at 1:2000 for 1 hour at RT. Counterstain: Mayer Hematoxylin (purple/blue) nuclear stain at 200 µl for 2 minutes at RT. Magnification: 40x.

<p>Immunohistochemistry analysis using Mouse Anti-DNA Damage Monoclonal Antibody, Clone 15A3 (SMC-155). Tissue: backskin. Species: Mouse. Fixation: Bouin’s Fixative and paraffin-embedded. Primary Antibody: Mouse Anti-DNA Damage Monoclonal Antibody (SMC-155) at 1:100 for 1 hour at RT. Secondary Antibody: FITC Goat Anti-Mouse (green) at 1:50 for 1 hour at RT.</p>

Immunohistochemistry analysis using Mouse Anti-DNA Damage Monoclonal Antibody, Clone 15A3 (SMC-155). Tissue: backskin. Species: Mouse. Fixation: Bouin’s Fixative and paraffin-embedded. Primary Antibody: Mouse Anti-DNA Damage Monoclonal Antibody (SMC-155) at 1:100 for 1 hour at RT. Secondary Antibody: FITC Goat Anti-Mouse (green) at 1:50 for 1 hour at RT.

<p>Immunohistochemistry analysis using Mouse Anti-DNA Damage Monoclonal Antibody, Clone 15A3 (SMC-155). Tissue: Ischemic fresh brain tissue. Species: Rat. Primary Antibody: Mouse Anti-DNA Damage Monoclonal Antibody (SMC-155) at 1:1000 for 16 hours at RT. Secondary Antibody: Alexa Fluor 546 Goat Anti-mouse (Red) at 1:500 for 1 hour at RT. Localization: Cerebral Cortex. Courtesy of: Dr. Yi Yang, U. New Mexico.</p>

Immunohistochemistry analysis using Mouse Anti-DNA Damage Monoclonal Antibody, Clone 15A3 (SMC-155). Tissue: Ischemic fresh brain tissue. Species: Rat. Primary Antibody: Mouse Anti-DNA Damage Monoclonal Antibody (SMC-155) at 1:1000 for 16 hours at RT. Secondary Antibody: Alexa Fluor 546 Goat Anti-mouse (Red) at 1:500 for 1 hour at RT. Localization: Cerebral Cortex. Courtesy of: Dr. Yi Yang, U. New Mexico.

Product Citations (10)

Immunohistochemistry

Epilepsy: neuroinflammation, neurodegeneration, and APOE genotype.

Aboud, O., Mrak R.E., Boop F.A., and Griffin W.S.T. (2013) Acta Neuropathol Commun. 1(1):41.

PubMed ID: 24252240 Reactivity Human Applications: Immunohistochemistry

Immunocytochemistry/Immunofluorescence

17β-Estradiol enhances sulforaphane cardioprotection against oxidative stress.

Angeloni, C. et al. (2017) J Nutri Biochem. 42 (2017) 26-36.

PubMed ID: N/A Reactivity Rat Applications: Immunocytochemistry/Immunofluorescence

In vivo oxidative stress alters thiol redox status of peroxiredoxin 1 and 6 and impairs rat sperm quality.

Liu, Y. and O'Flaherty, C. (2016) Asian J Androl. 19(1):73-79.

PubMed ID: 26823067 Reactivity Rat Applications: Immunocytochemistry/Immunofluorescence

Neutrophil extracellular traps enriched in oxidized mitochondrial DNA are interferogenic and contribute to lupus-like disease.

Lood, C. et al. (2016) Nat Med. 22(2):146-53.

PubMed ID: 26779811 Reactivity Various species Applications: Immunocytochemistry/Immunofluorescence

Neutrophil extracellular traps enriched in oxidized mitochondrial DNA are interferogenic and contribute to lupus-like disease.

Lood, C. et al. (2016) Nat Med. 22(2):146-53.

PubMed ID: 26779811 Reactivity Various species Applications: Immunocytochemistry/Immunofluorescence

Neuroprotective effect of sulforaphane against methylglyoxal cytotoxicity.

Angeloni, C. et al. (2015) Chem Res Toxicol. 28(6):1234-45.

PubMed ID: 25933243 Reactivity Human Applications: Immunocytochemistry/Immunofluorescence

Advancing age increases sperm chromatin damage and impairs fertility in peroxiredoxin 6 null mice.

Ozkosem, B., Feinstein, S.I., Fisher, A.B. and O'Flaherty, C. (2015) Redox Biol. 5:15-23.

PubMed ID: 25796034 Reactivity Mouse Applications: Immunocytochemistry/Immunofluorescence

Absence of PEROXIREDOXIN 6 is Associated with Low Sperm Chromatin Quality and Subfertility in Mice Challenged with Oxidative Stress.

Ozkosem, B. and O'Flaherty, C. (2013) figshare.

PubMed ID: N/A Reactivity Mouse Applications: Immunocytochemistry/Immunofluorescence

Other Citations

Disruption of redox homeostasis leads to oxidative DNA damage in spermatocytes of Wolbachia-infected Drosophila simulans.

Brennan, L.J., Haukedal, J.A., Earle, J.C., Keddie, B. and Harris, H.L. (2012) Insect Mol Biol. 21 (5): 510-520.

PubMed ID: 22831171 Reactivity Aedes albopictus Applications: Dot Blot

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