Name: Stabiele isotoop in-vivo labeling voor massaspectrometrie-identificatie van vaderlijke metabolieten die tijdens de bevruchting van sperma naar eicel worden overgebracht (Video) | JoVE Journal
Uploaded: 2025-06-17T14:00:00.000+00:00
Duration: 5 min 55 s
Description: Leer stabiele isotooplabeling voor massaspectrometrie om vaderlijke metabolieten te identificeren die bij de bevruchting van sperma naar eicel worden overgebracht. Deze methode onthult de rol van vaderlijke bijdragen aan embryonale ontwikkeling.

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5 minutes

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<table><tbody><tr><td>2H2O gefilterd, 99,8 atoom % 2H</td><td>Sigma Aldrich</td><td>756822</td><td></td></tr><tr><td>Aceton</td><td>Sigma Aldrich</td><td>179124</td><td></td></tr><tr><td>Agilent&nbsp; SIL-8</td><td>Agilent (Santa Clara, CA, U.S.A.)</td><td></td><td></td></tr><tr><td>C16-Ceramide-13C3-15N</td><td>LGC standard</td><td>TRC-C342770-25MG</td><td></td></tr><tr><td>dichloormethaan</td><td>Sigma Aldrich</td><td>34856</td><td></td></tr><tr><td>Diethylether&nbsp;</td><td>Sigma Aldrich</td><td>673811</td><td></td></tr><tr><td>ethanol</td><td>Sigma Aldrich</td><td>1,11,727</td><td></td></tr><tr><td>HCG (Human chorionic gonadotropin,</td><td>Sigma Aldrich</td><td>CG5-1VL</td><td></td></tr><tr><td>L-Lysine-&epsilon;-15N hydrochloride</td><td>Sigma Aldrich</td><td>608971</td><td></td></tr><tr><td>Mannelijke en vrouwelijke&nbsp; C57BL/6JOlaHsd muizen</td><td>Inotiv&nbsp;</td><td>C57BL/6JOlaHsd</td><td></td></tr><tr><td>methyl tert-butylether (MTBE)</td><td>Sigma Aldrich</td><td>306975</td><td></td></tr><tr><td>MetOH</td><td>Sigma Aldrich</td><td>1,06,035</td><td></td></tr><tr><td>N,O-Bis(trimethylsilyl(TMS) trifluoraceetamide (BSTFA)</td><td>Supelco</td><td>15238</td><td></td></tr><tr><td>PBS</td><td>Applichem</td><td>A0965-9010</td><td></td></tr><tr><td>PMSG (Pregnant mare&rsquo;s serum)</td><td>Lee Biosolutions</td><td>493-10</td><td></td></tr><tr><td>pyridine</td><td>Sigma Aldrich</td><td>360570</td><td></td></tr><tr><td>Shimadzu GCMS 2010plus&nbsp;</td><td>Shimadzu</td><td></td><td></td></tr><tr><td>Sitostanol-5,6,22,23-d4</td><td>Sigma Aldrich</td><td>680028</td><td></td></tr><tr><td>natriumtrifluoracetaat</td><td>Sigma Aldrich</td><td>1615138</td><td></td></tr><tr><td>SolariX XR 7T&nbsp;</td><td>Bruker Daltonics</td><td></td><td></td></tr><tr><td>TimsTOF Pro Q-TOF&nbsp;</td><td>Bruker Daltonics</td><td></td><td></td></tr><tr><td>U13C glucose&nbsp;</td><td>Sigma Aldrich</td><td>389374</td><td></td></tr><tr><td>Ultimate RS3000 UHPLC&nbsp;</td><td>Thermo Fisher Scientific</td><td></td><td></td></tr><tr><td>VanGuard CSHTM precolumn (5.0 &times; 2.1 mm; 1.7 &mu;m, 130 &Aring;)</td><td>Waters</td><td></td><td></td></tr></tbody></table>

stabiele isotoop in-vivo labeling voor massaspectrometrie-identificatie van vaderlijke metabolieten die tijdens de bevruchting van sperma naar eicel worden overgebracht

Stable Isotope In-Vivo Labeling for Mass-Spectrometry Identification of Paternal Metabolites Transferred from Sperm to Oocyte During Fertilization

Stabiele isotoop in-vivo labeling voor massaspectrometrie-identificatie van vaderlijke metabolieten die tijdens de bevruchting van sperma naar eicel worden overgebracht

Paternal contribution to embryo genetics has so far been limited to allelic sequences for decades. A decade of results suggests, instead, that epigenetic ...

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Biologie

Technieken voor Imaging Ca 2 + Signalering in Human Sperm

Stabiel isotopische profilering door middel van gaschromatografie massaspectrometrische analyse van intermediaire metabole flux wordt beschreven in de...

Stabiele Isotopen Profilering van Intermediair metabole flux in het ontwikkelen en volwassen stadium Caenorhabditis elegans</em

Dit protocol zal de extractie en analyse van vrije en geësterde bioactieve vetzuren uit cellen demonstreren. Vetzuren worden nauwkeurig gekwantificeerd met behulp van stabiele isotoopverdunning, chirale vloeistofchromatografie, electronenvangst-atmosferische chemische ionisatie-meervoudige reactiebewaking massaspectrometrie (SID-LC-ECAPCI-MRM/MS).

Cellulaire vetextractie voor gerichte stabiele isotopen Verdunning vloeistofchromatografie-massaspectrometrie Analyse

Bisulfiet mutagenese is de gouden standaard voor het analyseren van DNA-methylatie. Onze gewijzigde protocol zorgt voor een DNA-methylatie analyse op de single-cell niveau en is speciaal ontworpen voor individuele eicellen. Het kan ook worden gebruikt voor splitsing-embryo's.

Single Eicel bisulfiet Mutagenese

Proteomische analyse van elk celtype is sterk afhankelijk van zowel de zuiverheid als de pre-fractionering van het uitgangsmateriaal om het monster te de-complexificeren vóór vloeistofchromatografie-massaspectrometrie (MS). Door gebruik te maken van back-flushingtechnieken kunnen pure spermatozoa worden verkregen van knaagdieren. Na vertering kunnen fosfopeptiden worden verrijkt met behulp van TiO2.

Fosfopeptide Analyse van knaagdieren Epididymal Spermatozoa

We worden stappen beschreven waarmee snel in situ bemonstering van een klein gedeelte van een afzonderlijke cel met hoge precisie en minimale invasie met behulp van capillaire gebaseerde micro-sampling, ter vergemakkelijking van chemische karakterisering van een momentopname van de metabole activiteit in levende embryo's met behulp van een custom-built eencellige capillaire elektroforese en massaspectrometrie platform.

Microprobe capillaire elektroforese massaspectrometrie voor eencellige metabolomica in Live kikker (Xenopus laevis) embryo 's

Het vetzuur trofische marker aanpak, dat wil zeggen, de assimilatie van vetzuren als hele molecuul en transfer in consument weefsel met geen of een kleine wijziging, wordt gehinderd door lacunes in de kennis in de vetzuurstofwisseling van kleine bodem-ongewervelde dieren. Isotopologue profilering wordt geleverd als een waardevol instrument te ontwarren trofische interacties.

Profilering van vetzuur 13C Isotopologue geeft inzicht in de trofische Carbon overdracht en lipide metabolisme van ongewervelde consumenten

Hier beschrijven we een op massaspectrometrie gebaseerde proteomische karakterisering van cellijnen met bekende weefsellotgevallen in het gewervelde Xenopus laevis-embryo .

Cel-Lineage geleide massaspectrometrie proteomics in het zich ontwikkelende (kikker) embryo

Hier wordt een methode gepresenteerd voor het screenen van lipidestructuren door middel van stabiele isotopenlabeling door gebruik te maken van hun retentietijd, mobiliteit en fragmentatie.

De novo lipiden traceren met behulp van stabiele isotopenetikettering LC-TIMS-TOF MS/MS

Deze video toont de voorbereiding van een ultra-dunne matrix / analyt laag voor het analyseren van peptiden en eiwitten door Matrix-assisted laser desorptie ionisatie massaspectrometrie (MALDI-MS).

MALDI Monstervoorbereiding: de Ultra Thin Layer methode

Leer stabiele isotooplabeling voor massaspectrometrie om vaderlijke metabolieten te identificeren die bij de bevruchting van sperma naar eicel worden overgebracht. Deze methode onthult de rol van vaderlijke bijdragen aan embryonale ontwikkeling.

Stabiele isotoop in-vivo labeling voor massaspectrometrie-identificatie van vaderlijke metabolieten die tijdens de bevruchting van sperma naar eicel worden overgebracht (Video) | JoVE Journal

Stabiele isotoop in-vivo labeling voor massaspectrometrie-identificatie van vaderlijke metabolieten die tijdens de bevruchting van sperma naar eicel worden overgebracht

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