Name: Döllenme Sırasında Spermden Oosite Aktarılan Baba Metabolitlerinin Kütle Spektrometresi Tanımlaması için Kararlı İzotop In-Vivo Etiketleme (Video) | JoVE Journal
Uploaded: 2025-06-17T14:00:00.000+00:00
Duration: 5 min 55 s
Description: Keşfedin, kararlı izotop etiketleme ve kütle spektrometresi kullanarak döllenme sırasında spermden oosite aktarılan baba metabolitlerini. Bu yöntem erken embriyonik gelişimde epigenetik kalıtım mekanizmalarını anlamaya yardımcı olur.

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5 minutes

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<table><tbody><tr><td>2H2O filtrelenmiş, %99,8 atom 2H</td><td>Sigma Aldrich</td><td>756822</td><td></td></tr><tr><td>Aseton</td><td>Sigma Aldrich</td><td>179124</td><td></td></tr><tr><td>Agilent&nbsp; SIL-8</td><td>Agilent (Santa Clara, Kaliforniya, ABD)</td><td></td><td></td></tr><tr><td>C16-Seramid-13C3-15N</td><td>LGC standardı</td><td>TRC-C342770-25MG</td><td></td></tr><tr><td>diklorometan</td><td>Sigma Aldrich</td><td>34856</td><td></td></tr><tr><td>Dietil eter&nbsp;</td><td>Sigma Aldrich</td><td>673811</td><td></td></tr><tr><td>etanol</td><td>Sigma Aldrich</td><td>1,11,727</td><td></td></tr><tr><td>HCG (İnsan koryonik gonadotropini,</td><td>Sigma Aldrich</td><td>CG5-1VL Serisi</td><td></td></tr><tr><td>L-Lizin-&epsilon;-15N hidroklorür</td><td>Sigma Aldrich</td><td>608971</td><td></td></tr><tr><td>Erkek ve kadın&nbsp; C57BL/6JOlaHsd fareler</td><td>Inotiv&nbsp;</td><td>C57BL/6JOlaHsd</td><td></td></tr><tr><td>metil tert-bütil eter (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(trimetilsilil(TMS) trifloroasetamid (BSTFA)</td><td>Supelco</td><td>15238</td><td></td></tr><tr><td>PBS</td><td>Uygulama</td><td>A0965-9010 Serisi</td><td></td></tr><tr><td>PMSG (Hamile kısrak s serumu)</td><td>Lee Biyolojik Çözümler</td><td>493-10</td><td></td></tr><tr><td>piridin</td><td>Sigma Aldrich</td><td>360570</td><td></td></tr><tr><td>Shimadzu GCMS 2010plus&nbsp;</td><td>Şimadzu</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>Sodyum triflorasetat</td><td>Sigma Aldrich</td><td>1615138</td><td></td></tr><tr><td>SolariX XR 7T&nbsp;</td><td>Bruker Daltoniği</td><td></td><td></td></tr><tr><td>TimsTOF Pro Q-TOF&nbsp;</td><td>Bruker Daltoniği</td><td></td><td></td></tr><tr><td>U13C glukoz&nbsp;</td><td>Sigma Aldrich</td><td>389374</td><td></td></tr><tr><td>Ultimate RS3000 UHPLC&nbsp;</td><td>Thermo Fisher Bilimsel</td><td></td><td></td></tr><tr><td>VanGuard CSHTM ön sütunu (5.0 &kez; 2.1 mm; 1.7 &mu; m, 130 &Aring;)</td><td>Sular</td><td></td><td></td></tr></tbody></table>

döllenme sırasında spermden oosit'e aktarılan baba metabolitlerinin kütle spektrometresi tanımlaması için kararlı İzotop in-vivo etiketleme

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

Spermden oosite metabolit transferini izlemek için, ETİKETLİ ve ETİKETSİZ gruplardan sirelerle çiftleştirilmiş C57 BL/6J dişilerden izole edilen morulalar toplanır ve Kütle Spektrometresi tabanlı Metabolomikler ile analiz edilir. Metabolitler, adım 6-7'de tarif edildiği gibi ekstrakte edildi ve adım 8'de tarif edildiği gibi TMS ile türetildi. ETİKETLİ morulalar, ETİKETLENMEMİŞ morulalar, ETİKETLENMEMİŞ oositler, ETİKETLİ ve ETİKETLENMEMİŞ sperm hücrelerinin metabolomik ve lipidomik profillemesi yapıldı. Elde edilen profi...

Döllenme Sırasında Spermden Oosit'e Aktarılan Baba Metabolitlerinin Kütle Spektrometresi Tanımlaması için Kararlı İzotop In-Vivo Etiketleme

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

stable-isotope-vivo-labeling-for-mass-spectrometry-identification

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JoVE Journal

Biology

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Görüntüleme Ca Teknikleri 2 + İnsan Sperm sinyal

Kararlı izotop profil aracı metabolik akı gaz kromatografisi kütle spektrometrik analizi, nematod...

Kararlı İzotopik Profil geliştirilmesi ve Yetişkin Sahne Aracı Metabolik Akı Caenorhabditis elegans</em

Bu protokol, hücrelerden serbest ve esterlenmiş biyoaktif yağ asitlerinin ekstraksiyonunu ve analizini gösterecektir. Yağ asitleri, kararlı izotop seyreltme, kiral sıvı kromatografisi, elektron yakalama, atmosferik kimyasal iyonizasyon, çoklu reaksiyon izleme, kütle spektrometresi (SID-LC-ECAPCI-MRM/MS) kullanılarak doğru bir şekilde ölçülür.

Hücresel Lipid Ekstraksiyon Hedefli Kararlı İzotop Seyreltme Sıvı Kromatografi-Kütle Spektrometresi Analizi

Bisülfit mutagenez DNA metilasyon analizi için altın standarttır. Bizim modifiye protokolü tek hücre düzeyinde DNA metilasyon analizini sağlar ve özellikle bireysel oositler için tasarlanmıştır. Ayrıca, bölünme aşamalı embriyo için de kullanılabilir.

Tek Oosit Bisülfit Mutagenez

Herhangi bir hücre tipinin proteomik analizi, sıvı kromatografi kütle spektrometresinden (MS) önce numuneyi dekompleksten arındırmak için başlangıç materyalinin hem saflığına hem de ön fraksiyonlanmasına büyük ölçüde bağlıdır. Ters yıkama teknikleri kullanılarak, kemirgenlerden saf spermatozoa elde edilebilir. Sindirimi takiben, fosfopeptitler TiO2 kullanılarak zenginleştirilebilir.

Kemirgen Epididimal Spermatozoa&#39;lar phosphopeptide Analizi

Biz hızlı içinde in situ örnekleme yüksek kesinlik ve en az işgal kılcal tabanlı mikro-örnekleme, kimyasal bir anlık görüntüsünü metabolik aktivite karakterizasyonu kolaylaştırmak için kullanarak tek bir hücre bulunan küçük bir bölümünün etkinleştirmek adımları tarif canlı embriyolar bir ölçüye göre tek hücre Kapiler Elektroforez ve kütle spektrometresi platformu kullanarak.

Microprobe kılcal Elektroforez kütle spektrometresi canlı Kurbağa (Xenopus laevis) embriyo tek hücreli Metabolomics için

Yani, yağ asitleri asimilasyon tüm molekül ve transfer tüketici doku içine yağ asidi trofik marker yaklaşım değişiklik, engel olmayan ya da küçük küçük toprak omurgasızlar yağ asidi metabolizması bilgi boşlukları tarafından. Isotopologue profil oluşturma trofik etkileşimleri disentangle için değerli bir araç olarak sağlanır.

Yağ asidi 13C profil oluşturma Isotopologue trofik karbon Transfer ve Lipid metabolizması, omurgasız tüketicilerin sağlar

Burada, omurgalı Xenopus laevis embriyosunda bilinen doku kaderlerine sahip hücre soylarının kütle spektrometrisine dayalı proteomik karakterizasyonu açıklanmaktadır.

Gelişmekte Olan (Kurbağa) Embriyoda Hücre-Soy Kılavuzlu Kütle Spektrometrisi Proteomikleri

Burada sunulan, alıkonma sürelerini, hareketliliklerini ve parçalanmalarını kullanarak kararlı izotop etiketleme yoluyla lipid yapılarını taramak için bir yöntemdir.

Kararlı İzotop Etiketleme LC-TIMS-TOF MS/MS kullanarak de novo Lipidlerin İzlenmesi

Bu video Matriks Yardımlı Lazer Desorpsiyon İyonizasyon Kütle Spektrometresi (MALDI-MS) ile peptitler ve proteinlerin analizi için bir ultra-ince matris / analit bir tabaka hazırlık göstermektedir.

MALDI Numune Hazırlama: Ultra İnce Tabaka Yöntemi

Keşfedin, kararlı izotop etiketleme ve kütle spektrometresi kullanarak döllenme sırasında spermden oosite aktarılan baba metabolitlerini. Bu yöntem erken embriyonik gelişimde epigenetik kalıtım mekanizmalarını anlamaya yardımcı olur.

Döllenme Sırasında Spermden Oosite Aktarılan Baba Metabolitlerinin Kütle Spektrometresi Tanımlaması için Kararlı İzotop In-Vivo Etiketleme (Video) | JoVE Journal

Döllenme Sırasında Spermden Oosit'e Aktarılan Baba Metabolitlerinin Kütle Spektrometresi Tanımlaması için Kararlı İzotop In-Vivo Etiketleme

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