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Anti-HTT PHOSPHO Antibody
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|Huntingtin pS421 Antibody was prepared from whole rabbit serum produced by repeated immunizations with a synthetic peptide corresponding aa 416-424 of Human huntingtin protein.|
||ELISA: 1:10,000 - 1:40,000, WB: 1:500 - 1:3,000, IHC: 1:50 - 1:100
|0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2|
|Huntingtin (also known as Huntington's disease protein, Htt and HD protein) is the protein product of a disease gene linked to Huntington's disease, a neuro-degenerative disorder characterized by loss of striatal neurons. This may be caused by an expanded, unstable trinucleotide repeat in the huntingtin gene, which translates as a polyglutamine repeat in the protein product (see partial protein sequence below). The huntingtin gene locus is large, spanning 180 kb and consisting of 67 exons. It is expressed as 2 alternatively polyadenylated forms displaying different relative abundance in various fetal and adult tissues. The genetic defect leading to Huntington's disease may not necessarily eliminate transcription, but may confer a new property on the mRNA or alter the function of the protein. One candidate is the huntingtin-associated protein-1, highly expressed in brain, which has increased affinity for huntingtin protein with expanded polyglutamine repeats. Normal huntingtin protein shows a cytoplasmic localization. This protein is widely expressed with the highest level of expression in the brain (nerve fibers, varicosities, and nerve endings). In the brain, the regions where it can be mainly found are the cerebellar cortex, the neocortex, the striatum, and the hippocampal formation.
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Western blot analysis is shown using Affinity Purified anti-Huntingtin pS421 antibody to detect endogenous protein present in an unstimulated human PC-3 whole cell lysate (arrowhead). Comparison to a molecular weight marker indicates a band of ~190 kDa corresponding to truncated human Huntingtin protein. The blot was incubated with a 1:1,000 dilution of the antibody at room temperature followed by detection using standard techniques. Personal communication,
WB analysis after AKT and phosphatase treatment using Anti-Huntingtin pS421 antibody. In A) untreated N2A and 293A cells lysates were stained using anti-Huntingtin pS421 antibody. In B) staining after IP using a monoclonal antibody (Mab2166) followed by AKT treatment (to phosphorylate), along with untreated, and phosphatase (PP) treated (dephosphorylate) immunopreci-pitated Htt. In C) lysates are treated with AKT or PP.
Affinity Purified anti-Huntingtin pS421 antibody was used at a 1:100 dilution to detect phosphorylated Huntingtin by immunohistochemistry in human brain cerebellum. Positive cytoplasmic staining is observed in neurons. Tissue was formalin-fixed and paraffin embedded. Personal Communication,