[Novus] Exosome Isolation | 엑소좀 분리 컬럼, 비드, 침전용액 | 고마바이오텍

특징

• 엑소좀은 단백질, 지질, 핵산 등 다양한 세포 신호 물질을 포함하고 있는 세포외 소포체(Extracellular Vesicle; EV)의 한 종류입니다. 
• EV의 정확한 역할을 규명하기 위해서는 우선 생체액이나 세포배양 배지에서 EV를 분리해야 합니다.
• EV를 분리하는 데에는 침전법, 초고속원심분리법, 크로마토그래피, 면역친화 분리법 등이 있으며, 선택한 분리 방법에 따라 EV의 수율과 특이성이 결정됩니다. 
• NOVUS에서는 EV 분리 침전 용액, SEC 컬럼, 면역침강비드, 농축필터 등을 제공합니다. 
• 분리된 EV는 나노추적 분석(NTA), 단백질 프로파일링(WB, ELISA, Flow cytometry), 핵산 추출 등 다양한 후속 분석에 사용될 수 있습니다. 

• 세포 배양 배지처럼 희석된 샘플의 경우 EV를 분리하려면 더 빠르고 긴 원심 분리 과정을 거쳐야 합니다. 
• 소변의 경우 신장·방광·전립선 유래 EV를 얻는데 유용하지만 Tamm-Horsfall Protein (THP)가 많아 EV 탐지가 어려워질 수 있습니다. 
  
  

데이터

EV Precipitation Solution (Blood) [NBP3-11765] - Plasma - Exosomes isolated from plasma were resuspended in 100 ul of PBS and protein contents quantified by BCA assay. 60 ug of total protein contents were loaded with Laemmli Sample buffer 5X on acrylamide gel. WB performed using anti-ALIX antibody. Ultracentrifuged exosomes used as control (UC) Serum - Exosomes isolated from serum were resuspended in 24 ul of PBS 1X, and the entire amount was loaded with 6 ul of Laemmli Sample buffer 5X on acrylamide gel.

EV Precipitation Solution (Blood) [NBP3-11765] - Plasma and serum: After exosome isolation resuspend pellet in 100 ul of PBS 1X and load the entire amount in a well of an ELISA plate. Example reports CD9 expression in isolated exosomes from different volumes of plasma.

EV Precipitation Solution (Blood) [NBP3-11765] - Exosome pellet can be directly lysed with lysis buffers for nucleic acids extraction. If RNA extraction is performed by using Trizol or similar organic reagents we suggest resuspending exosomes in PBS, then to add Trizol into the mixture and to proceed with RNA purification.

EV SEC Columns [NBP3-11763] - Results and EV separation. EV SEC column was filled with 2 ml of medium from HCT116 cells, concentrated 10 folds by Tangential Flow Filtration (TFF)-EV concentrator (Catalog # NBP3-11762). 24 fractions (500 μl each one) have been collected and analyzed by ELISA assay (CD81 marker) and by BCA test for determining respectively vesicle and total protein content. EVs are eluted in fractions 8 - 12 (turnaround time approximately 15 min), whereas circulating proteins corresponded to the fractions 14 - 24.

EV SEC Columns [NBP3-11763] - Collected fractions were analyzed by NTA (Zetaview, Particle Metrix), ELISA and by BCA assay for determining EV and total protein content. EVs are eluted in fraction 2

CD9 Immunobeads for Exosome Isolation [NBP3-11768] - Anti-Alix WB analysis on exosomes immunocaptured with beads for overall (NBP3-11768) and for tumor-derived exosomes (NBP3-14798). WB shows the capture of exosomes only for cancer patients when beads for tumor-derived exosomes are used.image 1 of 2previousnextclose

CD63 Immunobeads for Exosome Isolation [NBP3-11769] - Alix expression by western blotting of exosomes captured on NBP3-11769 immunobeads from COLO1 cell supernatant vs isotype coupled beads