XF Glycolysis Stress Test Kit User Manual
Contents
1. IV Running the Glycolysis Stress Test Cel Seed Seed 24 hours pror io the inan XF cet culize melt pcena 7 Cnt piislsbtuentinipesiniopeien Myrning he XF Sensor Corrige The X Sens Cre must be ted priar 1o assay Piace the XF Sensor Cie on tap al the uy piste and plaea ina 37 C incubator Preparation ot Giyeolysis Stress Test Assay Medium Prepare the Test Medium trom Baze as o page 7 Prepare Compounds tor injection Traw DUSO are ene aliquot each o gueoe 2 54 Ogomyein 2 30 M at roem 1 Ata glucose ml 2am Svens Modum pe wart 7C ejesgitt a5 05 2 Final eencentratan ot guasa in assay wil be shore ioci 2 igni band on esl cl he n Opiimiatan Assays n page 9 See bie below rens E ET m m E osa in E sus vana Final eoncentraton oi 2 OGin assay 0020 Loss injection Poris te prepared compounds nio e spprgrt injection per and place in 37 C cron WIRED UR Perform the Medium Exchange O SQ ol coire medum each wet 2o Hsec imes with OD X Sec Tes Goin Astay Medium Ad 5104A to2. ard cet paie asdisev Data Analysis and interpretation The XF Gyon Sese Te scare cut graphe sies edi he pines sed oars arecornmandaton aa e calleedng derat aid io actieven M Figure 4 cela ers seoda at eril c 10 00 celu Intis casa the seedgderaty y tre is 20 000 cet el shau ba balanced qui belges Obame enli plnia a rrtcopa balira and atar ta arees bo seeding norm and cel Cel which are ovi or wel may lord Sehr a tgh degree variston barawer Te apima co seni be deemed sd on eol erra o Sanding density tat tate mitin te ess porien ot he graph 2 contrmaton sopra cel merpeiogy and seeing NOTE Ae mambar presant in mu af pyas pirunsonmenariirumn inside WR EUR Oligomycin Titration coreaneaton at Oigemyincanvary dca between derent types tue enserevrtl rear Therefor Pe opel Olgemeinconcrrlen odit deemed on any new cni p belare nunning he Gic Sess Test Seed 24 hors pror toe atsay in an XF cet and erani meti stan nas teen cf ri a resa tamem amets Far aquia oaa tla tla pera Hydrating he XF Sensor Cartridge The
3. x jr Prior priar te in an XF cl cue mpi glace ina muanbar et ota prement i aeg a neni e AF sana rete done get r Hydrating the XF Sensor Cartridge The XF Sensor must be hydrated priar to the assay the XF Sanar Cartridge on tapal the uy pile in a 97 COs tar a minmin 12 hee Preparation ot Giycoiysis Optimization Medium Prepare e Stress Opin Medium e Assay Bae edm at descrie on he Medium Exchange O 1 5044 ol the cure medium Korm 2 Pinse cel two imes ot XF Tesi Optimization Medum Pisce he pistein RP tor ane houe prer to a p Burning the with the XF Giyeolysis Test Software Ap pene XF satiare ihe Seshrze Apps down menu choose Start Aap on the Optimization Pte bution neate the cel seed deni tor esch eoum any veli come and ern ring he 70 Piace the estratan piate mih aded Vihen prampted replace aiy piste OK en Consina Cptrizaton Assay wit now na on XF
4. between Gy ie Capacty and Gaii ate deines yai WRITER 7 Glycolysis Stress Test Contents The XF Gies Tet contains areg reagents te un 6 tul XF assay plater Seahorse Glucose Scion vien 25M eln Seatorse gane powder t emply alo recondiues aliqui 1 valot powder 6 ernpty or reote Sestorse DSO Feqired not provided in e Seshora Bioscience o OD Serien vastei 2 piste ar Corte inaister setio 37 E XF Gian Svens Test rcormmended astay medium as on page 7 7 x casera toe kw phosphate DMEM Ss 05000 3 Res e g Soma P0290 Guesa teig Soma Gans 23030 Il Preparing the Components of the Assay Preparation of Assay Glycolysis Stress Test Base Medium Prepare the Day Prior to tne Assay DAEM D 5020 Sigma powder m Pesci rectore 1 ane batte ot Signa in 990ml al TC grade water pro 17 C 2 NaC tara fnat Van jette pio Tas 8 ard C Li stable at d G orales 3 monk Glycolysis Stress Test Optimization Medium Prepare the Day o e Preparation 100m ses Pare 100m al Gel Test Base Medium in apporte core 25M
5. lt estasion o protons heresi mediam As WERE EUR When phosgeryionis denpledby ATP ORgonyri el may increase erae ci lei iii ATP ad enr meri e resa shown Vu M efa This elevated rale o ipai riore io Pe Gli Capacty a iie Fata 1 XF Pet rd ard comprehersve moo saec 1e key parameters ol icai Gc and Reserve CHR abaone cl cos ts toic results Hom chr metabole pathways occuring in ecl Tree irat aigomyein and 2 Deory D 206 are injected consecutively 3 ECAR measurements aher each jection Tort bjectoniassurtogconceraon of cose 10 taten up by e ard eatabalized trough he pce patay to lacate prodi ATP The eetuin poor o the surrounding edm produces a agi in ECAR response repone as te rate a cis cl Nar under a basal ATP shins ha anergy produson to gyooiysis sth te subsequent increasa in ECAR capacty al he Th tinal njeten 205 4 glucose anslag wich gei tough comgeliv bn t guess e Srst enzyme n he pee Tte resuling decreasein ECAA conim that e ECAR e exprimens hue o
6. Glycolysis Stress Test Kit User Manual For use with the XF24 Analyzer Wade Turre UE Eik Preface Copyright Customer support Phone 00016710822 Option Tw Emab sapetgeesneseio cam WURDEN UA Table of Contents Introduction Preparing the Components ol the Assay Optimization Assays Funning the GlycolsisStess Tas Analyzing XF Giycolysis Stress Tes Data shoe ioci Introduction Overview rough tese enzyra oxidation and educta reactions Cale take up or ter ervronment sch as guess and proceesPem blat o generate resi in Pe usen prodr sch lt Ward CO irto the eara envronment ATP Tre Jose mestre there energy poing paw on aei etumeotriehaacs Tho rate ot OCA is Wheat etaceiar rough te poto repens genis micehorarespirton playin human and deesse Changes in dotes agro are crier cler immunalogy more varelatanal medene Assay Principle orae proshoraton are e energy producing ales ecl posesa me aiia evt dpa boten ese two processes ung elc ten convened ta iactata n e cytoplasm ar 2 in minori Glucose conversion 1o cie
7. ion tral concentration ol Tnt Glycolysis Stress Test Assay Medium repre the Dey o the Asay Preparation bm ueri assay 1 100m al Gel Base Meum in gore core 2 Wam to37 C andadas he pH 107 359 0 05 Reagent Preparation NOTE Waar chm Igea nen nanding een man bs partons Cuni ag ey Rt enum Bene XF yc Siess KA rom a eorr and alow the Olgomycin nd DO il we corake Preparation ot Olgomycin Aliquots T Sain donni a stack a coger appro 5 second perta geni 2 Besospendponderin Yol DMSO Translar M0 te vach algul vial Stare vie net nodad a 209 at re suspension on he side o te XF Swess Test KA WIRED UR Y x Preparation at 2 Deor D Glucose y NOTE praeter 00 ste may pem argu bonte but 2 25 005 jar rapa adjust volame o 10m wn Swess Tes Asray 3 eaen siqui Str visls rol needs medi 20C Record tne date re on the side o te XF Tesi Trawa tesh andi 2G foreach warm to room temperat betore Optimization Assays Cell Number Titration Metabole rates OCR ard ECAR can vary irat betwen derent cel ypes clu cordia may reque ned dereitas es
8. XF Sensor Cavidge must be hydrated priar to assay the XF Sanar Cartridge en tapal the uy piste and place in a 97C incubator COs tar aminin d 12 hi Day or A Preparation at Optimization Medium Persson pito Medium o the Qa Medium at cnn Perorm he Medium Exchange BOTE rb napa pts HO oe d 1 Sag ol medum fom each wel 2o Hsec imes it Gies Optimisation Mestan pre wed io C seh wel tar a 72 we Pre 1 cor CO rore houe prer to he EE z x Prepare Oligomycin Titrations tor Injection rM alga hors Veo are onde place coge Burning the mith he XF Strese Tet Jp 1 Opan the Ecole Inthe Saharze Apps doun XE e Start Ane bution Rur Optimization Piste buon Sec trom cop down meru seeding n tne soper eid pmi anaele oK 70 Pace the and wti injectar perts on eig vay Were reise the ps mih 1e cet pite 1 When therin i ovar ow the prompt in e cote ce piste ard dacara
9. sen wel tar a stole ar Sta 2 itne Seahorse Apps dra ren chaose 3 Ck be Sr Aap tuton Pea Stress buon B Enter tne orator each group n the ppt and create piste layout Wnen nished pr layout are anew Siart bunan to begin Tae x Piace tne and wih aded M Msn hetero clc deny V Analyzing the XF Glycolysis Stress Test Data Gic Sirens Sar App delivers scorso uomen eig keyparameters oi cli uneton hcl Gneci Capaciy nd uote Reserve Upon XF Gips Swess Ted tab is present ented Svens Test EE in tabular tom to MOT ehunga we mae in she anay mate re apor uing e Fe Da Tan aa Ap t aa ma rania aa a
Download Pdf Manuals
Related Search