Abstract
The DNA cytosine deaminase APOBEC3B (A3B) is normally an antiviral factor in the innate immune response. However, A3B has been implicated in cancer mutagenesis, particularly in solid tumors of the bladder, breast, cervix, head/neck, and lung. Here, we report data on the generation and characterization of a rabbit monoclonal antibody (mAb) for human A3B. One mAb, 5210-87-13, demonstrates utility in multiple applications, including ELISA, immunoblot, immunofluorescence microscopy, and immunohistochemistry. In head-to-head tests with commercial reagents, 5210-87-13 was the only rabbit monoclonal suitable for detecting native A3B and for immunohistochemical quantification of A3B in tumor tissues. This novel mAb has the potential to enable a wide range of fundamental and clinical studies on A3B in human biology and disease.
Highlights
The APOBEC3 family of single-stranded DNA cytosine deaminases constitutes a vital arm of the innate immune response, which serves to restrict the replication of viruses and transposable elements [1,2,3,4,5]
To determine whether the 5210-87-13 monoclonal antibody (mAb) recognizes nuclearHeLa cells were transfected with an A3B-eGFP expression construct and subjected to immunofluorescent microscopy (IF) microscopy
We report the creation and validation of a rabbit mAb, 5210-87-13, which binds to the human DNA cytosine deaminase A3B
Summary
The APOBEC3 family of single-stranded DNA cytosine deaminases constitutes a vital arm of the innate immune response, which serves to restrict the replication of viruses and transposable elements [1,2,3,4,5]. Ongoing research by many groups is likely to clarify the relative contributions of each APOBEC enzyme in the mutagenesis of different tumor types, and new technologies and reagents including validated monoclonal antibodies (mAbs) will have essential roles in this process. Following overnight incubation with primary antibody, sections were rinsed in TBST for 5 min, and completely covered with post-primary rabbit anti-mouse IgG The slides were drip-dried, transferred to TBST for 5 min, and incubated with anti-rabbit poly-HRP-IgG
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