Immunostaining Antibody Eluent

 

Khoom npe	Cat. Tsis muaj.	Spec..
Immunostaining Antibody Eluent	G1266	30 ml ib

 

 

Cov khoom no yog ua hauj lwm rau cov elution ntawm thawj thiab theem nrab cov tshuaj tiv thaiv thaum lub sij hawm immunofluorescence (IF), immunohistochemistry (IHC), thiab immunocytochemistry (ICC) immunostaining ntawm cov ntaub so ntswg thiab hlwb. Qhov no Immunostaining antibody eluent muaj qhov zoo ntawm kev ua haujlwm yooj yim, mob me me thiab muaj zog elution nyhuv. Nws tuaj yeem ua tiav cov tshuaj tiv thaiv thawj thiab theem nrab yam tsis muaj kev puas tsuaj rau morphology thiab cov qauv ntawm cov qauv. Nws yog tshwj xeeb tshaj yog haum rau cov antibody elusion nyob rau hauv lub TSA immunofluorescence multi-label sim ntawm paraffin seem, khov seem, cell slides, cell smear, thiab lwm yam ntawm lub hlwb, pob txha, tawv nqaij, aortic valve, thiab lwm yam. Cov non-covalently khi cov thawj antibody. thiab lub hom phiaj protein, cov tshuaj tiv thaiv thib ob thiab thawj cov tshuaj tiv thaiv kab mob tau cuam tshuam thiab eluted kom tshem tawm cov cawv ntawm cov tshuaj tiv thaiv tom ntej.

 

 

2 ~ 8 degree cia thiab thauj, siv tau rau 6 lub hlis.

 

 

Tus naj npawb	Cheebtsam	G 1266-30 mL
G1266	Immunostaining antibody eluent	30 ml ib
Phau ntawv		1pc ib

 

 

Cov khoom no yog npaj txhij-rau-siv thiab tsis tas yuav dilution. Ua ntej siv, tshem tawm cov tshuaj tiv thaiv kab mob los ntawm lub tub yees thiab rov qab mus rau chav tsev kub ua ntej ntxiv me me ntawm cov tshuaj tiv thaiv kab mob eluent los npog cov qauv, ua kom sov hauv chav sov li 5min, tom qab ntawd muab pov tseg cov eluent, ntxiv cov eluent txaus kom npog tag nrho cov ntaub so ntswg. , incubate ntawm 37 degree rau 30min, tso lub swb nyob rau hauv TBST thiab ntxuav nws ntawm lub rooj co rau 3 zaug, rau 5min txhua zaus.

Daim ntawv thov ntawm cov tshuaj tiv thaiv kab mob eluent tau piav qhia ntxaws nrog cov piv txwv ntawm TSA fluorescent multi-labeling hauv paraffin seem:

1. Cov ntaub so ntswg tau dewaxed rau dej: Sequentially, cov seem tau muab tso rau hauv Eco-phooj ywg dewaxing tov I (G1128) rau 10 min - Eco-friendly dewaxing tov II (G1128) rau 10 min - Eco-friendly dewaxing tov III (G1128 ) rau 10 min - anhydrous ethanol Ⅰ rau 5 min - anhydrous ethanol Ⅱ rau 5 min - anhydrous ethanol Ⅲ rau 5 min - thiab ntxuav nrog dej distilled;

2. Antigen kho: Raws li cov ntaub so ntswg thiab hom tshuaj tiv thaiv kab mob, xaiv cov tshuaj kho antigen sib raug (G1219,pH6.0; G1207,pH8.0; G1218,pH9 .0) thiab kho hom kev kho antigen. Txoj kev kho dav dav yog cov cuab yeej kho antigen (Servicebio yav tom ntej cov khoom), microwave, siab siab thiab lwm yam kev kho kub. Thaum lub sij hawm tus txheej txheem no, cov tshuaj kho yuav tsum tau tiv thaiv los ntawm ntau dhau evaporation thiab yuav tsum tsis txhob qhuav. Tom qab ntuj txias, cov swb tau muab tso rau hauv PBS (G2156) thiab ntxuav los ntawm kev co ntawm lub pendulum rooj (SYC-Z100) rau 3 zaug, 5 feeb txhua zaus.

3. Pap Pen kos lub voj voog thiab kaw nrog hydrogen peroxide: Tom qab cov ntu tau qhuav me ntsis, kos lub voj voog ncig cov ntaub so ntswg nrog Pap cwj mem (G6100), uas feem ntau yog li 3 hli ntawm cov ntaub so ntswg. Tom qab lub voj voog tiav lawm, ntxuav lawv nrog dej ntshiab (G4701) lossis PBS (G2156) tom qab lub voj voog qhuav, thiab tom qab ntawd muab cov ntu tso rau hauv 3% hydrogen peroxide tov thiab incubate ntawm chav tsev kub rau 25 min thiaj li yuav thaiv cov endogenous peroxidase. . Tom qab kaw, tus swb tau muab tso rau hauv PBS (G2156) thiab ntxuav los ntawm co ntawm lub pendulum rooj (SYC-Z100) rau 3 zaug, 5 feeb txhua zaus.

4. Serum blocking: Tom qab cov seem tau qhuav me ntsis, 3% BSA lossis cov kua dej ntxiv rau hauv cov ntaub so ntswg thiab kaw hauv chav sov li 30 min, thiab cov tshuaj tiv thaiv thaiv tau txiav txim siab raws li hom tshuaj tiv thaiv thawj thiab theem nrab.

5. Ntxiv thawj cov tshuaj tiv thaiv kab mob: Maj mam co tawm cov tshuaj sealing, ntxiv cov tshuaj tiv thaiv thawj zaug rau hauv ib qho kev faib ua feem rau cov hlais, muab cov hlais tiaj tus rau hauv lub thawv ntub dej (SIB-20F), incubate ntawm 4 degree thaum hmo ntuj . (Ntxiv cov dej me me rau hauv qab ntawm lub thawv ntub dej kom tsis txhob muaj cov tshuaj tiv thaiv evaporating)

6. Ntxiv cov khoom siv HRP uas tau sau cov tshuaj tiv thaiv kab mob thib ob: Cov swb tau muab tso rau hauv PBS (G2156) thiab ntxuav ntawm lub pendulum shaker (SYC-Z100) rau 3 zaug, 5 feeb txhua zaus. Tom qab cov hlais tau qhuav me ntsis, cov ntaub so ntswg coated ntawm cov tshuaj tiv thaiv kab mob thib ob uas sau nrog HRP ntawm cov kab sib txuas ntawm cov tshuaj tiv thaiv thawj zaug tau ntxiv rau lub nplhaib thiab incubated ntawm chav tsev kub rau 50 min.

7. Ntxiv iF488-TSA: Cov swb tau muab tso rau hauv PBS (G2156) thiab ntxuav los ntawm co ntawm lub pendulum rooj (SYC-Z100) rau 3 zaug, 5 feeb txhua zaus. Tom qab cov hlais tau qhuav me ntsis, cov tshuaj npaj iF488-TSA (G1231) ua haujlwm kua tau ntxiv rau lub nplhaib thiab incubated rau 10 min ntawm chav tsev kub kom deb ntawm lub teeb. Tom qab incubation, cov swb tau muab tso rau hauv TBST (G2150) thiab ntxuav ntawm lub pendulum shaker (SYC-Z100) rau 3 zaug, 5 feeb txhua zaus.

8. Antibody eluent kev kho mob: Tom qab cov seem tau qhuav me ntsis, cov tshuaj tiv thaiv eluant rov qab los rau hauv chav tsev kub tau kis mus rau tag nrho cov ntaub so ntswg. Tom qab incubating nyob rau hauv chav tsev kub rau 5 feeb, cov tshuaj tiv thaiv eluant raug tshem tawm, thiab ib tug txaus npaum li cas ntawm cov tshuaj tiv thaiv eluant tau ntxiv dua kom npog tag nrho cov ntaub so ntswg. Tom qab incubating ntawm 37 degree rau 30 feeb, tus swb tau muab tso rau hauv TBST (G2150) thiab ntxuav ntawm lub pendulum rooj (SYC-Z100) rau 3 zaug. 5 feeb txhua lub sijhawm.

9. Serum blocking: Tom qab cov seem tau qhuav me ntsis, 3% BSA los yog cov dej ntshiab ntxiv rau hauv cov ntaub so ntswg thiab kaw hauv chav sov li 30 min, thiab cov tshuaj thaiv cov tshuaj tiv thaiv tau txiav txim siab raws li hom tshuaj tiv thaiv thawj thiab theem nrab.

10. Ntxiv cov tshuaj tiv thaiv thib ob: Maj mam co tawm cov tshuaj sib khi, ntxiv cov tshuaj tiv thaiv thawj zaug rau hauv ib qho kev faib ua feem rau cov hlais, muab cov hlais rau hauv lub thawv ntub dej (SIB-20F), incubate ntawm 4 degree thaum hmo ntuj . (Ntxiv cov dej me me rau hauv qab ntawm lub thawv ntub dej kom tsis txhob muaj cov tshuaj tiv thaiv evaporating)

11. Ntxiv cov khoom siv HRP uas tau sau cov tshuaj tiv thaiv kab mob thib ob: Cov swb tau muab tso rau hauv PBS (G2156) thiab ntxuav ntawm lub pendulum shaker (SYC-Z100) rau 3 zaug, 5 feeb txhua zaus. Tom qab cov hlais tau qhuav me ntsis, cov ntaub so ntswg coated ntawm cov tshuaj tiv thaiv kab mob thib ob uas sau nrog HRP ntawm cov kab sib txuas ntawm cov tshuaj tiv thaiv thawj zaug tau ntxiv rau lub nplhaib thiab incubated ntawm chav tsev kub rau 50 min.

12. Ntxiv iF555-TSA: Cov swb tau muab tso rau hauv PBS (G2156) thiab ntxuav los ntawm co ntawm lub pendulum rooj (SYC-Z100) rau 3 zaug, 5 feeb txhua zaus. Tom qab cov hlais tau qhuav me ntsis, cov kua ua haujlwm iF555-TSA (G1233) tau ntxiv rau lub nplhaib thiab incubated rau 10 min ntawm chav tsev kub kom deb ntawm lub teeb. Tom qab incubation, cov swb tau muab tso rau hauv TBST (G2150) thiab ntxuav ntawm lub pendulum shaker (SYC-Z100) rau 3 zaug, 5 feeb txhua zaus.

13. Antibody eluent kev kho mob: Tom qab cov seem tau qhuav me ntsis, cov tshuaj tiv thaiv eluant rov qab los rau hauv chav tsev kub tau kis mus rau tag nrho cov ntaub so ntswg. Tom qab incubating nyob rau hauv chav tsev kub rau 5 feeb, cov tshuaj tiv thaiv eluant raug tshem tawm, thiab ib tug txaus npaum li cas ntawm cov tshuaj tiv thaiv eluant tau ntxiv dua kom npog tag nrho cov ntaub so ntswg. Tom qab incubating ntawm 37 degree rau 30 feeb, tus swb tau muab tso rau hauv TBST (G2150) thiab ntxuav ntawm lub pendulum rooj (SYC-Z100) rau 3 zaug. 5 feeb txhua lub sijhawm.

Faj seeb: Cov kauj ruam 4-7 yog ib puag ncig ntawm cov txheej txheem tshuaj tiv thaiv kab mob, txhua puag ncig ntawm daim ntawv lo tshuaj tiv thaiv tsuas yog yuav tsum tau hloov TSA reagent nrog cov tshuaj fluorescein sib txawv (iF647-TSA G1232; iF594-TSA G1242 iF440-TSA G1250; G1252; iF750-TSA G1258). Tom qab ib puag ncig ntawm kev sau tshuaj tiv thaiv kab mob, ua rau cov tshuaj tiv thaiv kab mob eluent ntawm 37 degree rau 30 min, tshem tawm cov tshuaj tiv thaiv thawj thiab theem nrab uas tau sau rau hauv qhov puag ncig no, thiab tom qab ntawd txuas ntxiv mus ntxiv ntawm cov tshuaj tiv thaiv kab mob. Nyob ntawm tus naj npawb ntawm cov tshuaj tiv thaiv kab mob fluorescent multi-labeled, rov ua cov kauj ruam 4-7 kom txog thaum tag nrho cov tshuaj tiv thaiv kab mob ua tiav.

14. DAPI restaining nuclei: Tom qab cov seem tau qhuav me ntsis, DAPI dye tov (G1012) ntxiv rau lub voj voog thiab incubated rau 10 min ntawm chav tsev kub kom deb ntawm lub teeb.

15. Autofluorescence quenching: Cov swb tau muab tso rau hauv PBS (G2156) thiab ntxuav los ntawm kev co ntawm lub pendulum rooj (SYC-Z100) rau 3 zaug, 5 feeb txhua zaus. Tom qab cov ntu tau qhuav me ntsis, cov ntaub so ntswg tus kheej-fluorescence quencher (G1221-2) tau ntxiv rau hauv lub nplhaib thiab incubated rau 5 min nyob rau hauv qhov tsaus ntuj nti, thiab ces yaug hauv qab dej rau 10 min.

16. Sealing: Cov seem yog me ntsis qhuav thiab kaw nrog cov tshuaj tiv thaiv fluorescence quenching sealing ntsiav tshuaj (G1401).

17. Kev yees duab microscopic: Cov seem tau saib nyob rau hauv lub fluorescence microscope thiab cov duab raug sau, los yog scanned nrog lub fluorescence scanner.

 

 

1. Cov nyhuv ntawm cov tshuaj tiv thaiv elution nyob ntawm ntu thickness, elution kub thiab lub sij hawm elution. 4 ~ 8 μm slices feem ntau yog kho nyob rau hauv ib lub thawv ntub dej ntawm 37 degree rau 30 min.

2. Txhawm rau kom ntseeg tau tias cov nyhuv antibody elution thiab fluorescent multi-label labeling, nws raug nquahu kom sau cov qauv membrane protein thiab cytoskeleton protein ua ntej, ces cytoplasmic protein, thiab thaum kawg nuclear protein.

3. Yog tias qee cov tshuaj tiv thaiv muaj zog affinity thiab tsis yooj yim rau elute tag nrho (xws li cov ntaub so ntswg hlwb GFAP), qhov kub thiab txias yuav nce mus rau 50 degree rau 30 min, los yog tus naj npawb ntawm elution tuaj yeem nce ntxiv, uas yog, tom qab ntxiv. antibody eluent rau cov ntaub so ntswg thiab incubating ntawm 37 degree rau 30 min, lub antibody eluent raug tshem tawm, thiab ib tug tshiab antibody eluent ntxiv, thiab lub incubation txuas ntxiv ntawm 37 degree rau 30 min.

4. Qhov no antibody eluent muaj zog fluidity, thiab thaum cov zaj duab xis tsis nyob rau hauv horizontally, cov reagent yog ib qho yooj yim mus ntws tawm ntawm lub nplhaib, cuam tshuam cov nyhuv elution. Nws yog ib qho tsim nyog yuav tsum tau them sai sai rau cov ntu tso ncaj nraim thaum lub sijhawm ua haujlwm.

5. Hnav hnab looj tes, daim npog qhov ncauj thiab lub tsho (lab) kom tsis txhob kov cov tawv nqaij thiab qhov muag. Thaum muaj kev sib cuag nrog cov chaw rhiab heev, yaug tam sim ntawd nrog dej ntau.

6. Lub titer ntawm cov reagent dhau hnub tas sij hawm yuav txo qis, yog li thov siv nws nyob rau hnub tas sij hawm ntawm cov reagent.

 

Rau Kev Tshawb Fawb Siv Tsuas!

 

 

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