HEK 293 cells were transfected using Xfect (Takara Bio, Mountain View, CA) to express HA-EVL and myc-cortactin. 48 hours post-transfection cells were prepared with NP40-CHAPS buffer. Immunoprecipitation was performed using Pierce anti-HA magnetic beads (Thermo Fisher Scientific, Waltham, USA). Proteins immunoprecipitates were eluted in LDS buffer (Thermo Fisher Scientific, Waltham, USA) and analyzed using antibodies against HA- and Myc-tag (Cell Signaling Technology, Danvers, USA), profilin-1 (Abcam, Cambridge, MA, USA) and profilin-2 (MilliporeSigma, St. Louis, MO, USA).
Ha and myc tag
Manufactured by Cell Signaling Technology
Sourced in United States
HA- and Myc-tag are protein tags that are commonly used in molecular biology and biochemistry research. They are short peptide sequences that can be attached to a protein of interest, allowing for the detection and purification of the tagged protein. The HA-tag is derived from the human influenza hemagglutinin protein, while the Myc-tag is derived from the c-Myc protein. These tags are frequently employed in experiments involving protein expression, localization, and interactions.
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2 protocols using ha and myc tag
1
Coimmunoprecipitation of EVL and Cortactin
2
Coimmunoprecipitation of EVL and Cortactin
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HEK 293 cells were transfected using Xfect (Takara Bio, Mountain View, CA) to express HA-EVL and myc-cortactin. 48 hours post-transfection cells were prepared with NP40-CHAPS buffer. Immunoprecipitation was performed using Pierce anti-HA magnetic beads (Thermo Fisher Scientific, Waltham, USA). Proteins immunoprecipitates were eluted in LDS buffer (Thermo Fisher Scientific, Waltham, USA) and analyzed using antibodies against HA- and Myc-tag (Cell Signaling Technology, Danvers, USA), profilin-1 (Abcam, Cambridge, MA, USA) and profilin-2 (MilliporeSigma, St. Louis, MO, USA).
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