Western blot analysis of Phospho-Htt (pSer434) was performed on 293T cells transfected with Htt and treated with PPI inhibitor and/or CDK5 to increase phosphorylation of Htt at Ser434. Lysates were loaded onto a 4-20% Tris-HCl polyacrylamide gel, transferred to a PVDF membrane and blocked with 5% BSA/TBST for at least 1 hour. The membrane was probed with a Phospho-Htt (pSer434) polyclonal antibody (Product # PA1-088) at a dilution of 1:500 overnight at 4°C on a rocking platform, washed in TBS-0.1%Tween-20 and probed with a goat anti-rabbit IgG-HRP secondary antibody (Product # 31460) at a dilution of 1:30,000 for at least 1 hour. Chemiluminescent detection was performed using SuperSignal West Pico (Product # 34087).
|Tested species reactivity||Human, Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic phosphopeptide conjugated to KLH corresponding to GGSSC(pS)PVLSRK (427-438) of Human HTT|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Tested Applications||Dilution *|
|Immunofluorescence (IF)||1:100 - 1:200|
|Immunohistochemistry (IHC)||1:100 - 1:200|
|Western Blot (WB)||1:250 - 1:500|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
PA1-088 has been successfully used in immunofluorescence, immunohistochemistry, and Western blot applications on human and mouse samples.
Huntingtin is a disease gene linked to Huntington's disease, a neurodegenerative disorder characterized by loss of striatal neurons. This is thought to be caused by an expanded, unstable trinucleotide repeat in the huntingtin gene, which translates as a polyglutamine repeat in the protein product. The huntingtin locus is large, spanning 180 kb and consisting of 67 exons. The huntingtin gene is widely expressed and is required for normal development. It is expressed as 2 alternatively polyadenylated forms displaying different relative abundance in various fetal and adult tissues. The larger transcript is approximately 13.7 kb and is expressed predominantly in adult and fetal brain whereas the smaller transcript of approximately 10.3 kb is more widely expressed. 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.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.