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anti-DNA mouse monoclonal, AC-30-10, liquid, purified, sample

Immunofluorescence analysis of HeLa cells with anti-DNA antibody. Fixation was performed using 3% paraformaldehyde for 10 min at RT. Cells were blocked with 5% BSA in PBST (PBS + 0.1% Tween 20) for 1 h at RT and permeabilized with 0.3% Triton-X 100 in PBS for 10 min at RT. The primary antibody anti-DNA mouse monoclonal, AC-30-10 (Cat. No. 690014) was diluted in blocking buffer (antibody concentration 0.5 µg/ml) and incubated over-night at 4°C. The secondary antibody goat anti-mouse IgM Alexa488 was also diluted in blocking buffer (antibody concentration 2.5 µg/ml) and incubated for 30 min at 37°C and 30 min at RT. Nuclear DNA was stained with DAPI in blue.

Liquid

Lyophilized

Liquid

Cat. No. 690014S

Prices excl. VAT

Available, delivery time: 1-7 days

€96.00*

Datasheet
Key Features
  • Mouse monoclonal
  • Suitable for Dot Blots, ICC/IF and IHC
  • Reacts with all species
  • Isotype: IgM
Product description
Quantity 200 µl
Antibody Type Monoclonal
Host Mouse
Isotype IgM
Conjugate Unconjugated
Application Dot blot, ICC/IF, IHC
Purification Size exclusion chromatography
Reactivity All species
Storage 2-8°C
Intended use Research use only
Clone AC-30-10
Immunogen Double- and single-stranded DNA
Concentration 100 µg/ml (20 µg)
Formulation PBS pH 7.4 with 0.09% sodium azide and 0.5% BSA
Applications
Tested applications Tested dilutions
Immunocytochemistry (ICC)/ Immunofluorescence (IF) 1:200 (0.5 µg/ml)
Immunohistochemistry (IHC) - frozen 1:10 (10 µg/ml)
Immunohistochemistry (IHC) - paraffin 1:10 (10 µg/ml, microwave treatment recommended)
Dot Blot Assay dependent (on nitrocellulose membrane, after baking at 70°C)
Background


AC-30-10 stains DNA present in the nuclei and the mitochondria of cells and tissues. It is also a reliable positive control in assays for detection of DNA antibodies, especially for the detection of autoantibodies in autoimmune diseases. Due to the high sensitivity it can also be used for the detection of mycoplasma contamination in cell cultures.
Binding characteristics: reacts with all forms of native and denatured DNA; reactive also with lambda DNA and synthetic DNA.
Tested cultured cell lines: XLKE-A6 (Xenopus), RVF-SMC, PTK-2, HeLa.

References/Publications (26)
Publication
Species
Application
Publication Paffett, M. L. et al. Interactomic analysis reveals a homeostatic role for the HIV restriction factor TRIM5α in mitophagy., Cell Rep 39, 110797, (2022).
Species human
Application ICC-IF
Publication Tanaka, K. et al. Mammalian BCAS3 and C16orf70 associate with the phagophore assembly site in response to selective and non-selective autophagy., Autophagy 17, 2011-2036, (2021)
Species human
Application ICC-IF
Publication Xu, J. et al. Synergistic effects of autophagy/mitophagy inhibitors and magnolol promote apoptosis and antitumor efficacy., Acta Pharm Sin B 11, 3966-3982, (2021).
Species human
Application ICC-IF
Publication Piro-Mégy, C. et al. Dominant mutations in mtDNA maintenance gene SSBP1 cause optic atrophy and foveopathy. J.Clin.Invest. 130, 143-156 (2020)
Species human
Application ICC
Publication Qin, J. et al. ER-mitochondria contacts promote mtDNA nucleoids active transportation via mitochondrial dynamic tubulation. Nat. Commun. 11, (2020).
Species monkey
Application IHC-IF
Publication Bailey, L. et al. PrimPol is required for the maintenance of efficient nuclear and mitochondrial DNA replication in human cells. Nucleic.Acids.Res. , (2019)
Species human
Application ICC-IF
Publication Ramos, E. et al. Mitochondrial fusion is required for regulation of mitochondrial DNA replication. PLoS.Genet. 15, e1008085 (2019)
Species mouse
Application ICC-IF
Publication Zhou, W. et al. PD-linked CHCHD2 mutations impair CHCHD10 and MICOS complex leading to mitochondria dysfunction. Hum Mol Genet. 28, 1100-1116(2019).
Species human
Application ICC/IF
Publication Hensen, F. et al. Mitochondrial RNA granules are critically dependent on mtDNA replication factors Twinkle and mtSSB. Nucleic.Acids.Res. , (2019)
Species human
Application ICC-IF
Publication Heinz, K. et al. Peripheral re-localization of constitutive heterochromatin advances its replication timing and impairs maintenance of silencing marks. Nucleic.Acids.Res. 46, 6112-6128 (2018).
Species mouse
Application ICC-IF
Publication Torralba, D. et al. Priming of dendritic cells by DNA-containing extracellular vesicles from activated T cells through antigen-driven contacts. Nat.Commun. 9, 2658 (2018).
Species human
Application ICC-IF,FACS
Publication Riley, J. et al. Mitochondrial inner membrane permeabilisation enables mtDNA release during apoptosis. EMBO.J. 37,- (2018).
Species mouse
Application ICC-IF
Publication Nicholls, T. et al. Topoisomerase 3α Is Required for Decatenation and Segregation of Human mtDNA. Mol.Cell. 69, 9-23.e6 (2018)
Species human
Application ICC-IF
Publication Wang, L. et al. PTEN-L is a novel protein phosphatase for ubiquitin dephosphorylation to inhibit PINK1-Parkin-mediated mitophagy. Cell.Res. 28, 787-802 (2018)
Species human
Application ICC-IF
Publication Dhir, A. et al. Mitochondrial double-stranded RNA triggers antiviral signalling in humans. Nature. 560, 238-242 (2018)
Species human
Application ICC-IF
Publication Dong, L. F. et al. Horizontal transfer of whole mitochondria restores tumorigenic potential in mitochondrial DNA-deficient cancer cells. Elife 6, (2017).
Species mouse
Application ICC-IF
Publication Vo, M. T. et al. Tristetraprolin inhibits mitochondrial function through suppression of α-Synuclein expression in cancer cells. Oncotarget 8, 41903–41920 (2017).
Species human
Application IHC-IF
Publication Lazarou, M. et al. The ubiquitin kinase PINK1 recruits autophagy receptors to induce mitophagy. Nature 524, 309-314 (2015).
Species human
Application ICC-IF
Publication Wong, Y. C. & Holzbaur, E. L. F. Optineurin is an autophagy receptor for damaged mitochondria in parkin-mediated mitophagy that is disrupted by an ALS-linked mutation. Proc. Natl. Acad. Sci. U. S. A. 111, E4439-E4448 (2014).
Species human
Application ICC-IF
Publication Kukat, C. et al. Super-resolution microscopy reveals that mammalian mitochondrial nucleoids have a uniform size and frequently contain a single copy of mtDNA. Proc. Natl. Acad. Sci. U. S. A. 108, 13534-13539 (2011).
Species human
Application ICC-IF
Publication Wang, Y. & Bogenhagen, D. F. Human Mitochondrial DNA Nucleoids Are Linked to Protein Folding Machinery and Metabolic Enzymes at the Mitochondrial Inner Membrane S. J. Biol. Chem. 281, 25791-25802 (2006).
Species human
Application ICC-IF
Publication Legros, F., Malka, F., Frachon, P., Lombes, A. & Rojo, M. Organization and dynamics of human mitochondrial DNA. J. Cell Sci. 117, 2653-2662 (2004).
Species human
Application ICC-IF
Publication Scheer, U. Structure of lampbrush chromosome loops during different states of transcriptional activity as visualized in the presence of physiological salt concentrations. Biol. Cell 59, 33-42 (1987).
Species Triturus
Application IF
Publication Scheer, U. et al. High sensitivity immunolocalization of double and single-stranded DNA by a monoclonal antibody. Eur. J. Cell Biol. 43, 358-71 (1987).
Species mouse,rat,rat,xenopus,Crithidia cuciliae,Triturus,chicken,bovine,M13 single and double stranded,recombinant plasmid DNA
Application IHC (frozen),IF,ICC-IF,IEM,dot blot
Publication Scheer, U., Hansmann, P., Falk, H. & Sitte, P. Ultrastructural localization of DNA in two Cryptomonas species by use of a monoclonal DNA-antibody. J. Cell Biol. 42, 152-160 (1986).
Species C. spec
Application IEM
Publication Benavente, R. & Krohne, G. Involvement of Nuclear Lamins in Postmitotic Reorganization of Chromatin As Demonstrated by Microinjection of Lamin Antibodies. J. Cell Biol. 103, 1847-1854 (1986).
Species rat
Application ICC-IF
Downloads
File
Category
Size
Filetype
File Immunofluorescence Protocol
Category Protocol
Size 178.25 KB
Filetype pdf
Datasheet
MSDS
FAQs
The concentration of unpurified supernatant, ascites, unpurified guinea pig serum and unpurified rabbit serum is not determined.
The concentration of purified antibodies is mentioned on the datasheet.
For prediluted antibodies the concentration may vary from lot to lot. The concentration of these antibodies is not mentioned on the datasheet and can be requested at support@progen.com.

Most of our purified mouse antibodies contain 0.5% BSA as stabilizer. If BSA was added to the antibody solution, it is stated in the datasheet.
The supernatant format contains FCS proteins from cell culture medium supplemented with FCS.
The serum antibodies contain other proteins present in serum.

Please find the solvent for reconstitution and the amount of liquid for reconstitution on the antibody datasheet. The buffer formulation after reconstitution is also mentioned in the datasheet.
PROGEN offers different formats for mouse monoclonal antibodies.  Different antibodies may not be offered in each format, the following are all possible formats:
  1. Supernatant and supernatant concentrate: This format contains hybridoma cell culture supernatant. The antibody is not purified and the antibody concentration is not determined. The antibody concentration may vary from lot to lot. Therefore we recommend to titrate the optimal concentration for the application used for each new lot.
  2. Lyophilized, purified: This format contains purified antibody in lyophilized form. The reconstitution of this antibody is described in the datasheet. The buffer composition after reconstitution is also mentioned on the datasheet. 
  3. Liquid, purified: This format contains purified antibody in liquid format. The concentration is mentioned on the datasheet. 
  4. Prediluted, purified: This format contains purified antibody in liquid format. Most antibodies in this format are diluted to be ready-to-use for IHC with standard tissue. But some antibodies of this format need further dilution for IHC. This is mentioned on the datasheet.

Lyophilized antibodies can be stored at 2-8°C until expiration.
Most of our liquid antibodies and reconstituted lyophilized antibodies may be stored for short term storage (up to 3 month) at 2-8°C. For long term storage we recommend to store the antibody at -20°C in aliquots. Please avoid freeze and thaw cycles.
Most of our conjugated antibodies should be stored at 2-8°C.
The individual storage conditions are mentioned on the datasheet.

The expiration date of our antibodies is indicated on the product label.
We offer bulk amounts starting from 1 mg for many of our mouse monoclonal or recombinant antibodies. Please contact us for further information or use the “bulk button” on the product page.
  • The antibody is very well suited for the detection of mitochondrial DNA; it is extremely sensitive and the signal of mitochondrial DNA is much stronger as mtDNA is not packed as densely as the DNA in the nucleus. The antibody stains the DNA in the nucleus weaker as the mitochondrial DNA.
  • For staining of the nucleus fixation with methanol/acetone (5 min in methanol (-20°C) and 5 min in acetone (-20°C)) is recommended.
  • The antibody is also suitable to stain bacterial DNA or synthetic single stranded DNA.

Most of our antibodies contain 0.09% sodium azide as preservative. If a preservative is added, it is mentioned in the datasheet.
The optimal antibody dilution for your specific protocol and application needs to be titrated in your lab with your equipment and sample. The optimal dilution may vary between protocols and samples. A good dilution for starting the titration is the dilution mentioned in the datasheet. If the sample needs a specific treatment (eg. Antigen retrieval for IHC on FFPE sections) this should also be mentioned on the antibody datasheet. 
PROGEN antibodies are shipped at ambient temperature. The antibodies are stable at ambient temperature for the shipment period. Please store the antibodies as indicated in the datasheet upon arrival.

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