Custom solutions & partnerships Custom antibody development and commercial partnerships to advance your diagnostic and therapeutic discovery. Keep up to date with the latest events Full event breakdown with abstracts, speakers, registration and more View global event calendar Mouse monoclonal [33D3] to 5-methylcytosine (5-mC) Suitable for: IHC-P, IP, Southern Blot, Flow Cyt, IHC-Fr Reacts with: Species independent Isotype: IgG1 Specificity Modified base 5-methylcytidine found in DNA of plants and vertebrates.Raised against the modified ribonucleoside. Specific for the presence of a methyl group on carbon 5 of the pyrimidine ring. General notes Storage in frost-free freezers is not recommended. Should this product contain a precipitate microcentrifugation before use.While older lots have performed well in ICC, we have received inconsistent results with the latest lots. Unfortunately, we can no longer guarantee this antibody for use in ICC.The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q As Storage instructions Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. Primary antibody notes Ab10805 recognises the modified base 5-methylcytidine found in DNA of plants and vertebrates. DNA methylation is a DNA modification process, which is involved in the control of gene expression. Reports suggest that in tumours, DNA is frequently globally hypomethylated compared to the DNA from normal tissue. The Abpromise guarantee Our Abpromise guarantee covers the use of ab10805 in the following tested applications. The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user. IHC-P (4) Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. Use at an assay dependent concentration. Use 10ul of working dilution to label 1000000 cells in 100ul. (see Habib, M. et al. (1999)).ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. IHC-Fr Use at an assay dependent concentration. Notes IHC-PUse at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. IPUse at an assay dependent concentration. Southern Blot1/200. Flow CytUse at an assay dependent concentration. Use 10ul of working dilution to label 1000000 cells in 100ul. (see Habib, M. et al. (1999)).ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. Form The native context of double-stranded DNA may obstruct antibody binding to 5-methylcytosine. For successful detection of 5-methylcytosine, we recommend that the DNA is denatured to make the nucleotides accessible for the antibody. Denaturing methods vary depending on each application. Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-5-methylcytosine (5-mC) antibody [33D3] (ab10805)This image is courtesy of an Abreview submitted by Ms Sara Maj Wätjen Hyldig ab10805 staining 5-Methyl Cytidine in pig embryo (15 to 17 days) tissue section by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue underwent fixation in paraformaldehyde, heat mediated antigen retrieval in Tris-EDTA buffer, permeabilization in Triton X-100 and blocking in 2% BSA for 10 minutes at 25 C. The primary antibody was diluted, 1/100 (PBS + 2% BSA) and incubated with sample for 1 hour at 25 C. An Alexa Fluor 488 conjugated donkey polyclonal to mouse at 1/250 dilution, was used as secondary.See Abreview Publishing research using ab10805? Please let us know so that we can cite the reference in this datasheet. ab10805 has been referenced in 120 publications. Zhu X et al. Ubiquitination-mediated degradation of TRDMT1 regulates homologous recombination and therapeutic response. NAR Cancer 3:zcab010 (2021).PubMed: 33778494 Chen X et al. DNA Hypermethylation Induced by L-Methionine Leads to Oligodendroglial and Myelin Deficits and Schizophrenia-Like Behaviors in Adolescent Mice. Front Neurosci 15:659853 (2021).PubMed: 33958986 Wolny E et al. Defining the Cell Wall, Cell Cycle and Chromatin Landmarks in the Responses of Brachypodium distachyon to Salinity. Int J Mol Sci 22:N/A (2021).PubMed: 33477958 Li M et al. Regulation of MYB by distal enhancer elements in human myeloid leukemia. Cell Death Dis 12:223 (2021).PubMed: 33637692 Biondi LR et al. Quantification of Global DNA Methylation in Canine Mammary Gland Tumors via Immunostaining of 5-Methylcytosine: Histopathological and Clinical Correlations. Front Vet Sci 8:628241 (2021).PubMed: 33718471 View all Publications for this product Blocking step 20% FCS, 1% BSA as blocking agent for 1 hour(s) and 0 minute(s) Concentration: 20% Temperature: 25 C Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) Concentration: 10% Temperature: 20 C Preparation Cell harvesting/tissue preparation method: Tissue were dissociated with collagenase. Obtained cells were filtered and fixed.Sample buffer: Cells were analysed in PBS ab10805 specifically binds to single modified cytosine so it should bind to 5-Methylcytosine in RNA as well. Read More Thank you for contacting us. While we have no tested the specificity for 5-hmC in house, customers have reported that this clone does not recognize 5-hmC. Unfortunately, we do not currently have any data regarding this reactivity. Read More Thank you for contacting us.There are many anti mouse secondary FITC conjugated antibodies available - they are all fine. Please buy the following antibody; https://www.abcam.com/goat-polyclonal-secondary-antibody-to-mouse-igg1-heavy-chain-fitc-pre-adsorbed-ab98692.htmlThis antibody is a preadsorbed one. Please click the link for more information; https://www.abcam.com/index.html?pageconfig=resource rid=12956I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.Use our products? Submit an Abreview. Earn rewards!https://www.abcam.com/abreviews Read More Thank you for contacting us.ab10805 should recognize single 5-methyl cytidine.I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information. Read MorePlease note: All products are FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES For licensing inquiries, please contact partnerships@abcam.com