5-METHYLCYTOSINE Antibody (OASA08969)

Catalog No: OASA08969 (Formerly GWB-9C7B77)

Size:100UG

Product Info

• Predicted Species Reactivity: All Species
• Product Format: Liquid. Phosphate buffered saline
• Clonality: Polyclonal
• Isotype: Polyclonal IgG
• Host: Sheep
• Conjugation: AF: Azide Free
• Application: Immunofluorescence|Immunohistochemistry-Frozen|Immunohistochemistry-Paraffin|Immunoprecipitation|Western blot
  Preservative Stabilisers: None present
  Antiserum Preparation: Antiserum to 5-MeC was raised by repeated immunisation of sheep with highly purified antigen. Purified IgG was prepared by affinity chromatography.
  Approx Protein Conc: IgG concentration 1.0mg/ml
  Buffer Solutions: Phosphate buffered saline
• Reconstitution and Storage: -20°C
• Immunogen: Keyhole limpet haemocyanin (KLH) conjugated 5-methylcytosine.
• Predicted Homology Based on Immunogen Sequence: Broad
• Concentration: 1 mg/ml
• Specificity: 5-METHYLCYTOSINE
• Application Info: Immunohistology - Frozen(1):    10ug/ml
  Immunohistology - Paraffin(2):    10ug/ml
  Immunoprecipitation:    25 - 100ug/ml
  Western Blotting:    1:1000
  (1) Special conditions app;y - see [*]
  (2) Special conditions apply - see [*]
• Protocol Information: Citation: 1: Hiura H, Sugawara A, Ogawa H, John RM, Miyauchi N, Miyanari Y, Horiike T, Li Y, Yaegashi N, Sasaki H, Kono T, Arima T. A tripartite paternally methylated region within the Gpr1-Zdbf2 imprinted domain on mouse chromosome 1 identified by meDIP-on-chip.Nucleic Acids Res. 2010 Aug;38(15):4929-45. Epub 2010 Apr 12. PubMed PMID: 20385583; PubMed Central PMCID: PMC2926594.
  Species: Mouse
  Experiment Name: MeDIP-on-chip analysis
  Experiment Background: Genomic imprinting describes the expression of a subset of mammalian genes from one parental chromosome. DNA methylation is acquired on the paternal allele at 4 DMRs and on the maternal allele at 18 DMRs. To identify known and novel DMRs Tiling array technology and chromatin immunoprecipitation (ChIP-on-chip) has been successfully applied to decipher chromatin structure with the methylated DNA binding column technique using the antibody against 5-methyl-cytosine (methylated-DNA immunoprecipitation; meDIP).
  Experimental Steps: 1. DNA extracted from PG- and AG-derived cells and mature sperm was fragmented to ~200–1000 bp by sonication.2. Immunoprecipitation was carried out using a specific antibody for 5-methyl-cytosine (AbD , Oxfordshire, UK).3. Input and bound DNA was amplified by GenomePlex Complete Whole Genome Amplification kit.4. The relative enrichment of DMRs was determined by sequence-specific real-time PCR analyses using a 7500 Real Time PCR System and SYBR Premix Ex Taq II.5. For the tiling arrays, input DNA was labeled with a cyan-3 dye and bound DNA was labeled with cyan-5.6. DNAs were hybridized to the mouse whole genome tiling array. 7. The methylated sequences were compared between PG- and AG-derived cells and sperm DNA using ChIP Analytics 1.3 software.
  Number Of Protocols: 1

Target Info

• Description of Target: SHEEP ANTI 5-METHYLCYTOSINE:Azide Free