Complete GPI anchor of Thy-1 required for recognition by monoclonal antibodies raised against membrane bound form (A) Mature Thy-1 detected by western blot in the CEs of NHLF and RFL-6 stably transfected with <t>pcDNA3.1/Zeo(+)</t> (pcDNA) or pcDNA3.1/Zeo (+) with a recombinant form of Thy-1 cloned in: WT THY1 (THY1), THY1 – GPI(DAF) (DAF), THY1 – GPI(TR3) (TR3), or FLAG – THY1 (FLAG). (B) Mature Thy-1 detected by western blot in CE containing FLAG – Thy-1 or CM containing PI-PLC released FLAG – Thy-1 or FLAG – sThy-1 using K117 and AS02. After stripping the membranes of antibodies for detecting mature Thy-1, FLAG was detected using M2.
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