Recombinant APOBEC3A (A3A) protein Catalog No: 81285 Quantity: 10 µg Expressed In: E. coli Concentration: 0.252 µg/µl Source: Human
Buffer Contents: Recombinant APOBEC3A is supplied in 50 mM Tris, pH 7.4, 50 mM NaCl, 10% glycerol, 1 mM beta-mercaptoethanol, and 0.01% Tween-20.
Background: Apolipoprotein B mRNA editing enzyme, catalytic polypeptide-like 3A, also known as APOBEC3A, or A3A is a gene of the APOBEC3 family found in humans, non-human primates, and some other mammals. It is a single-domain DNA cytidine deaminase with antiviral effects. While other members of the family such as APOBEC3G are believed to act by editing ssDNA by removing an amino group from cytosine in DNA, introducing a cytosine to uracil change which can ultimately lead to a cytosine to thymine mutation, one study suggests that APOBEC3A can inhibit parvoviruses by another mechanism. The cellular function of APOBEC3A is likely to be the destruction of foreign DNA through extensive deamination of cytosine.
Protein Details: Recombinant APOBEC3A was expressed in E.coli as the full length protein (accession number NP_663745) according to the protocol detailed in Schutksy, et al., Nucleic Acids Res. 2017 Jul 27. The molecular weight of rAPOBEC3A is 23 kDa.
Application Notes: Recombinant APOBEC3A is suitable for use in bisulfite-free whole genome sequencing of 5- hydroxymethylcytosine (e.g., ACE-Seq), cytosine deamination, enzyme kinetics, inhibitor screening and selectively profiling.
References: Nondestructive, base-resolution sequencing of 5-hydroxymethylcytosine using a DNA deaminase. Schutsky EK, DeNizio JE, Hu P, Liu MY, Nabel CS, Fabyanic EB, Hwang Y, Bushman FD, Wu H, Kohli RM. Nat Biotechnol. 2018 Oct 8 APOBEC3A efficiently deaminates methylated, but not TET-oxidized, cytosine bases in DNA. Schutsky EK, Nabel CS, Davis AKF, DeNizio JE, Kohli RM. Nucleic Acids Res. 2017 Jul 27
Storage and Guarantee: Recombinant proteins in solution are temperature sensitive and must be stored at -80°C to prevent degradation. Avoid repeated freeze/thaw cycles and keep on ice when not in storage. This product is guaranteed for 12 months from date of receipt.
This product is for research use only and is not for use in diagnostic procedures.
North America 877 222 9543 • Europe +32 (0)2 653 0001 • Japan +81 (0)3 5225 3638 • www.activemotif.com Recombinant APOBEC3A (A3A) SDS-PAGE gel Protein was run on a 10% SDS-PAGE gel and stained with Coomassie blue.
MW: 23 kDa
HPLC-based DNA cleavage assay for APOBEC3A activity. A single-stranded DNA oligonucleotide bearing one deoxycytidine is incubated with recombinant APOBEC3A, leading to deamination of deoxycytidine, a dC dU conversion. Incubation of dU bearing oligonucleotide with uracil-DNA glycosylase creates an abasic site that is then highly susceptible to base-mediated strand cleavage. (A.) Intact, deoxycytidine- containing substrate oligonucleotide with an HPLC retention time of 9 minutes. (B.) APOBEC3A-deaminated and base-cleaved ssDNA oligo. Cleavage products have retention times of 8.1 and 8.4 minutes.
North America 877 222 9543 • Europe +32 (0)2 653 0001 • Japan +81 (0)3 5225 3638 • www.activemotif.com