5-Carboxylcytosine Antibody - BSA Free

Immunocytochemistry/Immunofluorescence: 5-Carboxylcytosine Antibody [NBP2-59148] - 293T cells were transfected with either the mouse FLAG-tagged wild-type Tet1 (Tet1 CD) or the catalytically inactive FLAG-tagged C-terminal domain of Tet1 (Tet1 mCD) and stained with the antibody against 5-caC , diluted 1:500, and with an anti-FLAG antibody, followed by DAPI counterstaining.

Immunoprecipitation: 5-Carboxylcytosine Antibody [NBP2-59148] - Immunoprecipitation was performed with the antibody against 5-caC on 2 ug of J1 ES genomic DNA, spiked with 1 pg of a control DNA fragment (approximately 700 bp from the RFP (Ring finger protein) gene) containing different cytosine modifications. The mC and hmC control DNA was generated by PCR with the corresponding nucleotide. The caC control fragment was obtained by in vitro methylation using M.SssI methyltransferase followed by oxidation with purified Tet2. The IP'd DNA was subsequently anaysed by qPCR using primers specific for the control DNA fragments and for GAPDH, used as a negative control. Figure shows the enrichment calculated as the ratio of the recovery of the control DNA versus the recovery of the GAPDH negative control.

Dot Blot: 5-Carboxylcytosine Antibody [NBP2-59148] - Analysis was performed using synthetic oligonucleotides containing different modified C-bases (indicated in red). 125 and 25 ng of the respective oligo's were bound to a Streptavindin-coated multi-well plate. The antibody was used at a dilution of 1:1,000. The binding of antibody to the DNA was measured by ECL chemiluminescence. Figure shows a high specificity of the antibody for the carboxylated cytosine.

• Reactivity: Hu, Mu, All-NA
• Applications: DB, ICC/IF, IP, ICC/IF
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Format: BSA Free

5-Carboxylcytosine Antibody - BSA Free Summary

• Immunogen: The exact immunogen is propietary information.
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Purity: Protein A purified

Applications/Dilutions

• Dilutions:
  • Dot Blot 1:500 - 1:1000
  • Immunocytochemistry/ Immunofluorescence 1:500
  • Immunofluorescence 1:500
  • Immunoprecipitation 4 ug/IP

Packaging, Storage & Formulations

• Storage: Store at -20C. Avoid freeze-thaw cycles.
• Buffer: PBS, 30% Glycerol
• Preservative: 0.05% Sodium Azide
• Purity: Protein A purified

Alternate Names for 5-Carboxylcytosine Antibody - BSA Free

• 5-caC

Background

5-carboxylcytosine (5caC) is the final oxidized derivative of 5-methylcytosine (5mC). 5mC is oxidized to 5-hydroxymethylcytosine (5hmC) which is then oxidized to 5-formylcytosine (5fC) then 5caC. Each of these oxidation steps are catalyzed by the Ten-Eleven Translocation (TET) family of enzymes. 5fC can then be further oxidized to 5-carboxylcytosine (5caC) by TET. Both 5fC and 5caC can be converted to unmodified cytosine by Terminal deoxynucleotidyl transferase (TdT) or Thymine DNA glycosylase (TDG) by base excision repair.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.