MitoTracker Liab CMXRos

 

Khoom npe	Cat. Tsis muaj.	Spec.
Mito Tracker Liab CMXRos	G1723-50UG	50 ugj

 

 

MitoTracker Liab CMXRos (Mitochondrial Liab Fluorescent Probe) yog ib qho X-rosamine derivative nrog lub luj molecular ntawm 531.52, ib qho excitation wavelength ntawm 578 ± 3 nm thiab emission wavelength ntawm 599 ± 4 nm. Cov dyes tuaj yeem nkag mus rau cov hlwb ncaj qha thiab nrhiav tau ntawm mitochondria thiab tuaj yeem khi rau mitochondria. Lub hauv paus ntsiab lus ntawm kev sau npe tshwj xeeb ntawm mitochondria yog qhov kev sojntsuam lub cev pob txha nrog cov nqi zoo tuaj yeem nyob ntawm qhov tsis zoo them mitochondria, thiab chloromethyl pawg ntawm qhov kev sojntsuam tuaj yeem khi rau pawg dawb sulfhydryl hauv mitochondria, uas ua lub luag haujlwm ntawm fluorescent labeling. ; Lub cell mitochondria tuaj yeem sau npe yooj yim los ntawm incubating lawv nrog cov hlwb nyob rau qee lub sijhawm, thiab tom qab sau npe ntawm mitochondria, mitochondria tuaj yeem ua tiav ntxiv los ntawm kev txhim kho thiab perforation, uas tuaj yeem siv rau kev sim ntau daim ntawv lo tom qab. .

 

 

Nkoj nrog dej khov; Khaws ntawm -20 degree deb ntawm lub teeb, siv tau 12 lub hlis.

 

 

Cheebtsam	G1723-50UG
Mito Tracker Liab CMXRos	50 ugj
Phau ntawv	1pc ib

 

 

1. Assay ua hauj lwm daws npaj:

1.1. Cov khoom no yog nyob rau hauv daim ntawv ntawm cov hmoov qhuav, uas yuav tsum tau centrifuged luv luv ntawm kev kub ceev ua ntej siv los xyuas kom meej tias nws nyob rau hauv qab ntawm lub raj. Muab koj tus kheej qib cell DMSO.

1.2. Kev npaj cov tshuaj cia: Ntxiv 94 μL DMSO (qib cell) mus rau hauv lub raj, tshuab thiab sib tov zoo kom tag nrho yaj cov dye, uas yog hais tias, mitochondrial fluorescent dye cia tov nrog ib tug concentration ntawm 1 mM (molecular luj cov ntaub ntawv ntawm lub khoom daim ntawv lo) tau txais, thiab tom qab ntawd centrifuge luv luv ntawm kev kub ceev kom tsis txhob poob ntawm reagents dai ntawm phab ntsa. Khaws cov tshuaj cia ntawm -20 degree . Nws raug nquahu kom khaws cia rau hauv qhov me me thiab tsis txhob rov khov thiab thawing.

1.3. Kev npaj ntawm kev tshuaj ntsuam xyuas ua haujlwm: Yog tias qhov kev ntsuam xyuas tau ua tiav ncaj qha tom qab staining, dilute cov tshuaj cia rau hauv mitochondrial assay ua hauj lwm cov tshuaj ntawm qhov kawg concentration ntawm 0.1-0.25 μM nrog cov tshuaj lom neeg (Hanks, PBS) ) los yog tsis muaj ntshav basal nruab nrab; Yog hais tias tom qab ua haujlwm xws li kho thiab perforation yuav tsum tau tom qab staining, dilute cov tshuaj cia rau hauv mitochondrial assay ua hauj lwm kev daws teeb meem ntawm qhov kawg concentration ntawm 0.4-0.6 μM nrog lom tsis zoo (Hanks, PBS) los yog tsis muaj ntshav basal nruab nrab;

2. Cell staining (cov txheej txheem hauv qab no yog rau cov hlwb sib txuas; centrifugation yuav tsum tau muab tshem tawm ntawm tes pauv)

2.1. Cov hlwb tau cog rau ntawm daim phiaj kab lis kev cai ntawm tes los yog cov phiaj nce toj kom txog thaum lawv sib sau ua ke rau qee yam;

2.2. Lub cell kab lis kev cai nruab nrab raug tshem tawm thiab ntxuav 1-2 zaug nrog preheated tsis rau 3-5 min txhua zaus;

2.3. Ntxiv cov tshuaj preheated ua hauj lwm rau mitochondrial nrhiav thiab incubate ntawm 37 degree rau 30 min nyob rau hauv lub cell kab lis kev cai incubator (vim muaj ntau hom ntawm tes thiab lub xeev, lub incubation lub sij hawm yuav tsum tau kho rau qee yam);

2.4. Tshem tawm cov tshuaj incubation thiab ntxuav nrog buffer tov 2-3 zaug rau 3-5 min txhua zaus;

2.5. Ncaj nraim los yog nrog Anti-fade Mounting Medium (pom zoo G1401) cov yeeb yaj kiab raug kaw thiab pom nyob rau hauv lub tshuab hluav taws xob fluorescence (Ex=579} nm, Em=599 nm);

3. Cell fixation thiab perforation thiab lwm yam manipulations (yeem):

Nco tseg: Tom qab staining, cov khoom no tuaj yeem kho thiab perforated, tab sis cov teeb liab fluorescent yuav raug txo qis tom qab kho.

3.1. Cell fixation

3.1.1. Cells kho nrog mitochondrial fluorescent dye labeling staining thiab ntxuav yog ntxiv rau fixative (pom zoo G1101) thiab tsau rau 10 min ntawm chav tsev kub;

3.1.2. Tshem tawm cov fixative thiab ntxuav nrog tsis 2-3 zaug rau 3-5 min txhua zaus;

3.2. Cell perforation

3.2.1. Cov hlwb ruaj khov thiab ntxuav tau ntxiv nrog 0.2-0.5% Triton X-100 thiab perforated rau 10 min ntawm chav tsev kub;

3.2.2. Cov kua perforating raug tshem tawm thiab ntxuav nrog cov tshuaj tsis haum 2-3 zaug rau 3-5 min txhua zaus;

3.3. Cov hlwb raug stained thiab tom qab ntawd tsau thiab perforated rau kev ua, thiab tuaj yeem siv rau kev sim tom ntej xws li multilabeling.

 

 

1. Vim muaj ntau hom thiab lub xeev ntawm cov hlwb, kev ua haujlwm concentration ntawm cov dye thiab lub sijhawm incubation tuaj yeem hloov kho kom tsim nyog.

2. Cov zas xim yuav tsum tau siv rau mitochondrial localization nyob rau hauv lub xeev ntawm cov cell nyob, thiab cov hlwb tsis tuaj yeem kho ua ntej, txwv tsis pub nws yuav ua rau cov txiaj ntsig tsis zoo ntawm qhov tsis raug hauv zos. Kev kho thiab kev ua haujlwm tom ntej tuaj yeem ua tau tom qab staining ntawm cov cell nyob.

3. Thaum staining thiab ntxuav cov hlwb nyob, qhov kev ntsuam xyuas ua haujlwm thiab kev ntxuav tsis huv yuav tsum tau ua kom sov ua ntej ntawm 37 degree. Staining thiab ntxuav yog ua tiav los ntawm incubation nyob rau hauv lub incubator thiaj li tsis mus cuam tshuam lub morphology ntawm lub hlwb los ntawm qhov kub thiab txias stimulation.

4. Txhawm rau tiv thaiv fluorescence quenching, tag nrho cov txheej txheem staining yuav tsum tau ua kom deb ntawm lub teeb.

5. Kev sojntsuam tswv yuav tsum muab khaws cia rau hauv cov thawv cais kom tsis txhob rov ua kom khov thiab thawing. Kom tsis txhob pov tseg, cov tshuaj staining yog npaj raws li xav tau, npaj siv.

6. Rau koj txoj kev noj qab haus huv thiab kev nyab xeeb, thov hnav lub tsho (lab) thiab hnab looj tes thaum ua haujlwm.

 

Rau Kev Tshawb Fawb Siv Tsuas!

 

 

Cim npe nrov: mitotracker liab cmxros, Tuam Tshoj mitotracker liab cmxros manufacturers, lwm tus neeg, hoobkas