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Pulldown of structural proteins

Pulldown of structural proteins - Immunoprecipitation Forum

Pulldown of structural proteins - IP or Immunoprecipitation Forum.

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Old 02-22-2009, 06:26 AM
Pipette Filler
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Default Pulldown of structural proteins

Hello. I would appreciate if anyone can help me.

From the yeast 2 hybrid results, the protein of my interest seems to interact with structural proteins in muscle and heart. Because there is no cell line currently avalilable that express structural proteins abundantly, I am planning to clone my gene into a vector with histidine tag and use Pierce’s product “Pull-Down PolyHis Protein:Protein Interaction Kit (Catalog Number 21277)”. Here is what I am planning:

Bait: clone in the gene of my interest into pcDNA(tm) 3.1/V5-His (Invitrogen);, which attaches 6x His and V5 at the C-terminus, and overexpress it in HEK293T cells.

Prey: skeletal muscle from the thigh of a mouse. (tissue extract)

Does anyone try such an approach? Or any other approach?

Thank you very much for your help in advance.

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proteins , pulldown , structural

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