{[ promptMessage ]}

Bookmark it

{[ promptMessage ]}

BIO 102 Lecture 3 Notes

3 warm fus ed het erok ary ons t o 37 degrees f ix c

Info iconThis preview shows page 1. Sign up to view the full content.

View Full Document Right Arrow Icon
This is the end of the preview. Sign up to access the rest of the document.

Unformatted text preview: an heterok ary ons . 3. Warm fus ed het erok ary ons t o 37 degrees , f ix (c hemic ally k ill/ c ros s link and "f reez e" c ells ) s at v arious t imes aft er warming 4. Immunos tain f us ed, f ix ed c ells wit h ant i mous e and ant i human antibodies 5. Vis ualiz e dis tribut ion of mous e and human c ell s urf ac e antibodies as a func t ion of t ime af ter warming At 0 t ime, half the c ell has mous e membrane prot eins , half has human's . Stain c ell wit h f luores c ent ant ibodies , s o half and half are s tained… af ter 40 minut es , both are t otally mix ed. Why t emperature dependent ? At lower degrees , membrane is more rigid. Ant ibody or lec t in induc ed pat c hing and c apping of c ell s urfac e prot eins demons t rat es induc ed lat eral mov ement ; patc hing is ATP independent bec aus e all energy is built int o binding s it es , c apping requires ATP (t here is an underly ing c y tos k eleton and mot or proteins are try ing to driv e t hem int o an aggregate). Direc t v is ualiz ation of membrane protein mobilit y (or lac k t hereof ) by F R A P a n d F L I P A nal y s i s : met hods f or f luores c ent ly t agging int egral membrane proteins : a) monov alent fluores c ent lec t ins (nons pec if ic ) -- monov alent bec aus e more would driv e pat c hing b) f luores c ent ly t agged monov alent (FAB f ragment s ) ant ibodies t o s pec ific protein c ) t rans fec t c ells with c ons t ruc t enc oding (GFP-t agged membrane prot ein) and t hen p...
View Full Document

{[ snackBarMessage ]}

Ask a homework question - tutors are online