Liphetoho tsa Peptide
Peptide Biotinylation
Peptide Biotinylation ke eng?
Ho fumanoa ha li-peptide tse tšoailoeng ka nepo kapa li-peptide tsa biotinylated (mohlala. e nang le li-antibodies tse ngotsoeng)Karohano ea li-peptide tse nang le biotinylated kapa li-peptide tse tšoailoeng ho tse sa tšoauoang (mohlala. ka li-antibodies tse koetsoeng)
Li-tag e ka ba limolek'hule tse nyane tse kang biotin (e tlamang ka matla le e seng covalently ho streptavidin) kapa tatelano e kgutshwane ya peptide. Mehlala e hlahelletseng ka ho fetesisa ea tatellano ea peptide e sebelisoang joalo ka li-tag ke li-tag tsa epitope joalo ka folakha ea folakha., tag ea HA le tag ea Myc. Letšoao le sa tsoa ntlafatsoa ke tag ea ALFA peptide. Bakeng sa li-tag tsohle, li-antibodies li fumaneha khoebong. Sheba ka tlase bakeng sa tatellano e fapaneng.
Liphetoho tsa MOL li lula li kopanya li-peptide tse nang le li-tag tse fapaneng. Hangata li-tag li khomaretsoe ho N-terminus kapa C-terminus (ka lysine kapa cysteine), empa ka molao-motheo e ka behoa kae kapa kae. Li-tag tsohle li ka aroloa ho peptide ka mefuta e fapaneng e bitsoang li-linkers kapa limolek'hule tsa spacer tsa bolelele bo fapaneng le polarity.. Haeba o lakatsa, li-linkers li ka hlakoloa, mohlala. ka ho fokotsa litlamo tse hlokolosi tsa disulfide. Sheba lethathamo la li-linker tse fumanehang / spacer / PEGYlations. Mona ke setšoantšo sa li-tag tse kenyellelitsoeng khafetsa.
Li-peptide tsa Biotinylated le tse Tagged
Li-tag tsa Affinity tse kang biotin li ka sebelisoa bakeng sa
· Ho fumanoa ha li-peptide tse tšoailoeng ka nepo kapa li-peptide tse nang le biotinylated (mohlala. e nang le li-antibodies tse ngotsoeng)
·Karohano ea li-peptide tse nang le biotinylated kapa li-peptide tse tšoailoeng ho tse sa tšoauoang (mohlala. ka li-antibodies tse koetsoeng)
Li-tag e ka ba limolek'hule tse nyane tse kang biotin (e tlamang ka matla le e seng covalently ho streptavidin) kapa tatelano e kgutshwane ya peptide. Mehlala e hlahelletseng ka ho fetesisa ea tatellano ea peptide e sebelisoang joalo ka li-tag ke li-tag tsa epitope joalo ka folakha ea folakha., tag ea HA le tag ea Myc. Letšoao le sa tsoa ntlafatsoa ke tag ea ALFA peptide. Bakeng sa li-tag tsohle, li-antibodies li fumaneha khoebong. Sheba ka tlase bakeng sa tatellano e fapaneng.
Liphetoho tsa MOL li lula li kopanya li-peptide tse nang le li-tag tse fapaneng. Hangata li-tag li khomaretsoe ho N-terminus kapa C-terminus (ka lysine kapa cysteine), empa ka molao-motheo e ka behoa kae kapa kae. Li-tag tsohle li ka aroloa ho peptide ka mefuta e fapaneng e bitsoang li-linkers kapa limolek'hule tsa spacer tsa bolelele bo fapaneng le polarity.. Haeba o lakatsa, li-linkers li ka hlakoloa, mohlala. ka ho fokotsa litlamo tse hlokolosi tsa disulfide.
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Affinity Tag
|
Tatelano
|
Maikutlo
|
|---|---|---|
|
Letšoao la folakha |
DYKDDDDK |
Anti-folakha ea anti-folakha |
|
HA letsatsi |
YPYDVPDYA |
Anti HA antibody |
|
Nako e ngata |
EQKLISEEDL |
Anti Myc antibody |
|
CPP
|
Tatelano
|
Maikutlo
|
|---|---|---|
|
Tat (47-57) |
YGRKRRQRR |
Peptide e kenang liseleng |
|
Oligo-Arginine |
RRRRRRR |
Peptide e kenang liseleng |
MOL Changes e boetse e khona ho u fa mefuta e mengata e fapaneng ea li-labels tse amanang le li-peptide. Mehlala ke selelekela sa
- Azides (karabelo le alkines - tobetsa k'hemistri)
- Ho penta (karabelo le thiols, mohlala. Cys)
- Thiols (karabelo le maleimides)
- Cys(Pys/Npys) (karabelo le thiol ho etsa disulfides)
Litlhaloso tse hlahang
Myeloid special CD33 ea motho ke receptor e thibelang batho e nang le tšebetso e fapaneng ea ITIM ho hira phosphatase SHP-1 le SHP-2.
CD33 ke setho se ikhethileng sa myeloid sa lelapa la sialic-binding-binding 'me se hlahisoa haholo liseleng tsa myeloid progenitor empa maemong a tlase haholo liseleng tse arohaneng.. CD33 ea motho e na le masalla a mabeli a tyrosine sebakeng sa eona sa cytoplasmic (Y340 le Y358). Ha phosphorylated, Li-tyrosine tsena li ka sebetsa e le libaka tsa ho emisa phosphatase, SHP-1 le/kapa SHP-2, e nolofalletsang CD33 ho sebetsa joalo ka seamoheli sa inhibitory. Mona re bontša hore CD33 ke tyrosine phosphorylated ka pel'a phosphatase inhibitor., pervanadate, mme e thaotha SHP-1 le SHP-2. Lithuto tsa co-expression li fana ka maikutlo a hore Src-family kinase Lck e sebetsa ho phosphorylating Y340., empa eseng Y358, e fana ka maikutlo a hore masala ana a ka sebetsa molemong oa khetho ea limolek'hule tsa adaptara mme a na le mesebetsi e ikhethileng.. Tšehetso e eketsehileng bakeng sa ho fetana, empa e sa hlokeng letho, karolo ea Y340 le Y358 e tsoa lithutong tse tlamang peptide tse senotseng ho hiroa ha SHP-1 le SHP-2 ho Y340 empa ke SHP-2 ho Y358 feela.. Tlhahlobo e sebelisang liphetoho tse fetotsoeng tsa SHP-1 e bonts'itse hore ho tlama Y340 ea CD33 e ne e le haholo-holo sebakeng sa amino Src homology-2 sa SHP-1.. Bokhoni ba hore CD33 e sebetse joalo ka sehokelo sa inhibitory bo bonts'itsoe ke bokhoni ba eona ba ho theola taolo ea CD64-induced calcium mobilization ho U937.. Ho itšetleha ha thibelo ena ho SHP-1 ho ile ha bontšoa ka ho thibela litlamorao tsa CD33 tse nang le SHP-1 e mpe haholo.. Sephetho sena se fana ka maikutlo a hore CD33 ke mokelikeli o thibelang likokoana-hloko le hore SHP-1 phosphatase e na le karolo ea bohlokoa ho tsamaellaneng le tšebetso ea CD33.. Liphuputso tse ling li bohlokoa ho khetholla li-receptor(s) hore CD33 e thibela ho vivo le ts'ebetso ea eona ho nts'etsopele ea leloko la myeloid.
Litšebeletso tse Amanang tsa Phetoho ea Peptide
glycosylation ea peptide
glycosylation ea peptide
Peptide glycosylation ke phetoho e kopanetsoeng e ka ntlafatsang thepa ea physicochemical ea peptide.
Bala haholoanyanePhosphorylation ea peptide
Phosphorylation ea peptide
Phosphoralation e ka hlaha ka Serine (S, Ho ba), Threonine (T, Thr) le Tyrosine (Y, Tyr) liketane tse mahlakoreng ka ho theha bond ea phosphoester
Bala haholoanyaneLi-peptide tse tloaelehileng
Li-peptide tse tloaelehileng
Kenyelletso ea li-amino acid tse peli tseo e seng tsa tlhaho tse nang le α-methyl, lihlopha tsa α-alkenyl nakong ea motsoako o tiileng oa liketane tsa peptide.
Bala haholoanyanePeptide ea cyclic
Peptide ea cyclic
Peptide cyclization e ntlafatsa botsitso ba li-peptide (e amanang le li-analogue tsa bona tsa mela) 'me ke leano le tloaelehileng ho nts'etsopele ea peptide.
Bala haholoanyanephetoho ea peptide
Boiphihlelo bo pharalletseng ba phetoho ea peptide, ho fana ka mekhoa e mengata e sebetsang ea lipatlisiso tsa peptide.
