Anti-DNA/RNA Damage antibody [15A3] (ab62623)

Mouse monoclonal DNA/RNA Damage antibody [15A3]. Validated in ELISA, IHC, ICC/IF and tested in Species independent. Cited in 53 publication(s). Independently reviewed in 12 review(s).

Overview

  • Product name

    Anti-DNA/RNA Damage antibody [15A3]
    See all DNA/RNA Damage primary antibodies
  • Description

    Mouse monoclonal [15A3] to DNA/RNA Damage
  • Host species

    Mouse
  • Tested applications

    Suitable for: IHC-P, ELISA, ICC/IF, IHC-Fr, IHC-FoFrmore details
  • Species reactivity

    Reacts with: Species independent
  • Immunogen

    Chemical/ Small Molecule corresponding to DNA/RNA Damage. 8-hydroxy-guanosine-BSA and -casein conjugates.

  • Positive control

    • IHC-P: Mouse inflammed colon and backskin tissues. Fresh IHC: Ischemic rat brain tissue.
  • General notes

    Please see the protocol booklet link below for recommended IHC and ICC staining procedure

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.09% Sodium azide
    Constituents: PBS, 50% Glycerol
  • Concentration information loading...
  • Purity

    Protein G purified
  • Clonality

    Monoclonal
  • Clone number

    15A3
  • Isotype

    IgG2b
  • Light chain type

    kappa
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab62623 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IHC-P Use a concentration of 1 - 10 µg/ml.

Dilute antibody in PBS containing 0.3% Triton X-100, 0.08% sodium azide and 2% normal goat serum.

ELISA Use at an assay dependent concentration.
AP Use at an assay dependent concentration.
ICC/IF 1/500 - 1/1000.

PubMed 25479606

IHC-Fr Use at an assay dependent concentration.
IHC-FoFr Use at an assay dependent concentration. PubMed: 21824519

Target

  • Relevance

    In intact animals, lesions (adducts) excised from DNA are transported from the cell through the circulation and excreted in urine. In bacteria, DNA adducts are excreted directly into the medium. In either case, the adducts can be assayed as a measure of oxidative damage to DNA. In particular, Oxo-8-dG (8-Oxo-7,8-dihydro-2'-deoxyguanosine) serves as an excellent marker for DNA damage produced by oxidants because it represents one of the major products generated by a wide array of treatments associated with oxidant damage such as that produced by irradiation and various carcinogens and because it is implicated in spontaneous transversion mutagenesis. Oxo-8-Gua (8-oxo-7,8-dihydroguanine) is one of the most common DNA lesions resulting from reactive oxygen species and can result in a mismatched pairing with adenine resulting in G to T and C to A substitutions in the genome. In humans, it is primarily repaired by DNA glycosylase OGG1. It can be caused by ionizing radiation, in connection with oxidative metabolism. Oxo-8-G (8-oxo-7,8-dihydroguanosine) is classified as an oxidized ribonucleotide, and is primarily used in studies of oxidative RNA damage and associated RNA repair and RNA turnover mechanisms within the cell. In the cell, Oxo-8-G RNA lesions are formed by reaction with reactive oxygen species (ROS) generated either via normal oxidative metabolic processes, UV ionizing radiation, or exposure to oxidative agents. Oxidative RNA damage can lead to defects in protein synthesis, for example, decreased rates of protein synthesis and production of aggregated or truncated peptides, with important implications in aging and neurodegenerative disorders and artherosclerosis.
  • Alternative names

    • 7,8-Dihydro-8-oxo-2'-deoxyguanosine antibody
    • 7,8-Dihydro-8-oxodeoxyguanosine antibody
    • 7,8-Dihydro-8-oxoguanine antibody
    • 8-Hydroxy-2'-deoxyguanosine antibody
    • 8-Hydroxydeoxyguanosine antibody
    • 8-hydroxyguanine antibody
    • 8-OH-dG antibody
    • 8-Oxo-2'-deoxyguanosine antibody
    • 8-Oxo-7,8-dihydro-2'-deoxyguanosine (Oxo-8-dG) antibody
    • 8-Oxo-7,8-dihydrodeoxyguanosine antibody
    • 8-oxo-7,8-dihydroguanine (Oxo-8-Gua) antibody
    • 8-oxo-7,8-dihydroguanosine (Oxo-8-G) antibody
    • 8-Oxo-dG antibody
    • 8-Oxo-G antibody
    • 8-oxo-Gua antibody
    • 8-Oxo-guanosine antibody
    • 8-Oxo-riboguanosine antibody
    • 8-Oxoguanine antibody
    • OH8Gua antibody
    see all

Images

  • Bouin’s Fixative, paraffin-embedded mouse backskin tissue stained for DNA/RNA Damage using ab62623 (1 hr at RT) at 1/100 dilution in immunohistochemical analysis.

    Secondary Antibody: FITC Goat Anti-Mouse (green) at 1/50 for 1 hour at RT.

  • Mouse hepatocytes stained for DNA/RNA Damage (green) using ab62623 at 1/500 dilution in ICC/IF, followed by Alexa-Fluor®488 conjugated Goat Anti-Mouse IgG (H+L).

    See Abreview

  • Immunohistochemical analysis of murine brain tissue 24 hours after recirculation following ischemia. Staining using ab62623 at 1/1000 dilution. An AlexaFluor®488-conjugated anti-mouse IgG (1/500) was used as the secondary antibody.

  • Left panel: ab62623 (1/1000 for 16 hours at RT) staining in ischemic rat brain tissue (fresh samples).
    Center panel: DAPI staining
    Right panel: merged

    Secondary Antibody: Alexa Fluor®546 Goat Anti-mouse (Red) at 1/500 for 1 hour at RT.

  • Immunohistochemical analysis of PFA-fixed paraffin-embedded rat femural tissue, labeling with ab62623 at a dilution of 1/50 incubated for 13 hours at 4°C in 1% BSA in TBS. Heat mediated antigen retrival was performed via Tris-EDTA pH 9.0. Blocking was via ab93695 ABC kit incubated at 1% for 20 minutes at room temperature. A secondary was not used, but ab93695 detection kit was used for signal amplification.

    See Abreview

  • ab62623 staining in rat liver tissue section by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).

    Tissue underwent formaldehyde fixation before enzymatic antigen retrieval with Proteinase K solution and then blocking for 20 minutes at 37°C was performed. The primary antibody was diluted 1/4000 and incubated with sample for 2 hours at 37°C. A Biotin conjugated rabbit polyclonal to mouse IgG was used as secondary antibody at 1/200 dilution.

    See Abreview

  • Paraffin-embedded mouse inflammed colon tissue stained for DNA/RNA Damage using ab62623 at 1/1,000,000 dilution (12 hrs at 4°C) in immunohistochemical analysis.

    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.

References

This product has been referenced in:

  • Millett CE  et al. The Sex-specific Effects of LPS on Depressive-like Behavior and Oxidative Stress in the Hippocampus of the Mouse. Neuroscience 399:77-88 (2019). Read more (PubMed: 30553795) »
  • Kopp F  et al. PUMILIO hyperactivity drives premature aging of Norad-deficient mice. Elife 8:N/A (2019). Read more (PubMed: 30735131) »
See all 61 Publications for this product

Customer reviews and Q&As

1-10 of 20 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Lung)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM sodidum citrate buffer pH 6.0
Permeabilization
Yes - 0.5% Triton X 100
Specification
Lung
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Jun 07 2019

Application
Immunohistochemistry (Frozen sections)
Sample
Rat Tissue sections (liver)
Permeabilization
No
Specification
liver
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 22°C
Fixative
Acetone

Abcam user community

Verified customer

Submitted Feb 05 2018

Application
IHC - Wholemount
Sample
Mouse Tissue (intestine organoids)
Specification
intestine organoids

Abcam user community

Verified customer

Submitted Apr 11 2017

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (hepatocytes)
Permeabilization
Yes - 0.1% Triton X-100 in TBS for 5-10 minutes
Specification
hepatocytes
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Aug 09 2016

Application
Immunocytochemistry
Sample
Oncorhynchus mykiss Cell (Spermatozoa)
Permeabilization
Yes - 0,3%Triton-X100 in PBS1X
Specification
Spermatozoa
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 20% · Temperature: 4°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Oct 06 2015

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (Bone tissue (Femur))
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris-EDTA pH 9.0
Specification
Bone tissue (Femur)
Blocking step
Ab93695 ABC kit as blocking agent for 20 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative
Paraformaldehyde

Mr. Helder Fonseca

Verified customer

Submitted Sep 11 2015

Application
ELISA
Sample
Human Cell (immortalized proximal tubule HK2 cells)
Specification
immortalized proximal tubule HK2 cells
Blocking step
Odyssey Blocking Buffer as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 100% · Temperature: 24°C
Type
Other

Abcam user community

Verified customer

Submitted Jul 22 2015

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 20°C
Sample
Mouse Cell (Hepatocyte)
Specification
Hepatocyte
Permeabilization
No
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Jan 23 2014

Question
Answer

We added IHC of paraffin-embedded tissue to the applications section of the ab62623 datasheet based on reviews we received from customers. These are linked on the main online datasheet. Each customer used a different method for antigen retrieval. One used a heat-mediated retrieval with citrate buffer at pH 6.0 (probably simmering at 95-100C for 15-20 minutes, followed by 20 minutes of cooling), before staining mouse skin, and the other customer used a solution of proteinase K for 4 minutes on sections of rat liver. We do not have any data for mouse lung.

I suggest trying a heat retrieval with 10mM citrate buffer pH 6.0. Please let me know if you need more details about how to do the antigen retrieval.

Here are links to the reviews:

https://www.abcam.com/8-Hydroxyguanosine-antibody-15A3-ab62623/reviews/18245

https://www.abcam.com/8-Hydroxyguanosine-antibody-15A3-ab62623/reviews/15116

Read More
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (B16F10 melanoma)
Specification
B16F10 melanoma
Fixative
4% Formalin
Permeabilization
Yes - 0.1% SDS
Blocking step
BSA as blocking agent for 15 minute(s) · Concentration: 1% · Temperature: 25°C

Ms. Seontae Kim

Verified customer

Submitted Mar 15 2013

1-10 of 20 Abreviews or Q&A

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