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Recombinant APOBEC3A (A3A) protein

Expressed In

E. coli

Protein Species

Human

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Cat No : 81285 81285

Validation Data Gallery

  • Recombinant APOBEC3A (A3A) SDS-PAGE gel Protein was run on a 10% SDS-PAGE gel and stained with Coomassie blue. MW: 23 kDa

    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.

    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.

Product Information

Expressed In E. coli
Protein Species Human

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.

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.

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.

Storage

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.

Guarantee

This product is guaranteed for 6 months from date of receipt.

This product is for research use only and is not for use in diagnostic procedures.