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Western blot analysis of HRV3c was performed by loading 1ug of a control protein, either uncleaved (lane 1) or cleaved (lane 2) using the Pierce Human Rhinovirus 3C Protease (HRV 3C) (Product # 88946) per well onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with 5% BSA/TBST for at least 1 hour. The membrane was probed with an HRV3c polyclonal antibody (Product # PA1-118) at a dilution of 1:1000 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 1:15,000 for at least 1 hour. Chemiluminescent detection was performed using SuperSignal West Dura (Product # 34075).
|Tested species reactivity||Virus|
|Host / Isotype||Rabbit / IgG1|
|Immunogen||Synthetic peptide recognizing the cleavage site of HRV3c protease|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS with 1mg/ml BSA, 30% glycerol|
|Contains||0.05% sodium azide|
|Tested Applications||Dilution *|
|Immunoprecipitation (IP)||4 µg|
|Western Blot (WB)||1:1000-1:2000|
* 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.
PA1118 has been used successfully in Western Blot to detect HRV3C cleavage site (Leu-Glu-Val-Leu-Phe-Gln-Gly-Pro).
PA1118 has been used in Immunoprecipitation of an HRV3C cleavage site-containing protein from a whole Hela cell lysate.
Human rhinovirus 3C protease (HRV3C Protease) is a cysteine protease that recognizes the cleavage site of Leu-Glu-Val-Leu-Phe-Gln-Gly-Pro. It cleaves between Gln and Gly (independent of Pro). (HRV) 3C Protease is used to remove fusion tags from proteins with the HRV 3C cleavage sequence and is typically dual tagged for easy removal from the sample after cleavage.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.