Khoshnan A, Ko J, Patterson PH. Effects of intracellular expression of anti-huntingtin antibodies of various specificities on mutant huntingtin aggregation and toxicity. Proc Natl Acad Sci U S A. 2002 Jan 22;99(2):1002-7. PubMed Abstract

REAGENTS/MATERIAL:

Eight mAbs (MW1), (MW2), (MW3), (MW4), (MW5), (MW6) (MW7) and (MW8) that bind the epitopes polyglutamine (polyQ), polyproline (polyP), or the C terminus of exon 1 in huntingtin (htt) protein were developed. Of these MW1 and MW2, recognize the polyQ domain, and MW7 recognizes the polyP domains of htt exon 1. MW1 stains htt in the cytoplasm, MW2 displays vesicular staining of htt, and MW7 stains htt in the perinuclear region as well in the nuclear inclusions formed by mutant htt. To examine the effects of these mAbs on the biological activities of htt exon 1, we have cloned and expressed the antigen-binding domains of these three mAbs as single-chain variable region fragment Abs (scFvs). Expression of scFvs was examined by Western blot using an anti-Flag Ab (1:1000, Sigma) and for Co-localization rabbit (Vivian Hook) and anti-Flag (1:1000; Sigma).