Protocol for cell surface biotinylation of magnetic labeled and captured human peripheral blood mononuclear cells
Summary: Analysis of the surfaceome of a blood cell subset requires cell sorting, followed by surface protein enrichment. Here, we present a protocol combining magnetically activated cell sorting (MACS) and surface biotinylation of the target cell subset from human peripheral blood mononuclear cells...
| Main Authors: | , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2022-12-01
|
| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166722007432 |
| _version_ | 1811292657399365632 |
|---|---|
| author | Serife Ayaz-Guner Mustafa Burak Acar Dudu Boyvat Huseyin Guner Habibe Bozalan Melis Güzel Selin Kübra Yıldır Nilay Altınsoy Fatma Fındık Musa Karakükçü Servet Özcan |
| author_facet | Serife Ayaz-Guner Mustafa Burak Acar Dudu Boyvat Huseyin Guner Habibe Bozalan Melis Güzel Selin Kübra Yıldır Nilay Altınsoy Fatma Fındık Musa Karakükçü Servet Özcan |
| author_sort | Serife Ayaz-Guner |
| collection | DOAJ |
| description | Summary: Analysis of the surfaceome of a blood cell subset requires cell sorting, followed by surface protein enrichment. Here, we present a protocol combining magnetically activated cell sorting (MACS) and surface biotinylation of the target cell subset from human peripheral blood mononuclear cells (PBMCs). We describe the steps for isolating target cells and their in-column surface biotinylation, followed by isolation and mass spectrometry analysis of biotinylated proteins. The protocol enables in-column surface biotinylation of specific cell subsets with minimal membrane disruption. : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| first_indexed | 2024-04-13T04:48:37Z |
| format | Article |
| id | doaj.art-48e46a8c0a2f4aa19c68c51d71634648 |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-04-13T04:48:37Z |
| publishDate | 2022-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-48e46a8c0a2f4aa19c68c51d716346482022-12-22T03:01:45ZengElsevierSTAR Protocols2666-16672022-12-0134101863Protocol for cell surface biotinylation of magnetic labeled and captured human peripheral blood mononuclear cellsSerife Ayaz-Guner0Mustafa Burak Acar1Dudu Boyvat2Huseyin Guner3Habibe Bozalan4Melis Güzel5Selin Kübra Yıldır6Nilay Altınsoy7Fatma Fındık8Musa Karakükçü9Servet Özcan10Department of Molecular Biology and Genetics, Izmir Institute of Technology, Izmir 35430, Türkiye; Department of Molecular Biology and Genetics, Faculty of Life and Natural Science, Abdullah Gül University, Kayseri 38080, TürkiyeFaculty of Science, Department of Biology, Erciyes University, Kayseri 38039, Türkiye; Genome and Stem Cell Center (GENKÖK), Erciyes University, Kayseri 38039, TürkiyeBioengineering Program, Graduate School of Engineering and Science, Abdullah Gul University, Kayseri 38080, TürkiyeDepartment of Molecular Biology and Genetics, Faculty of Life and Natural Science, Abdullah Gül University, Kayseri 38080, TürkiyeGenome and Stem Cell Center (GENKÖK), Erciyes University, Kayseri 38039, Türkiye; Gevher Nesibe Genome and Stem Cell Institute, Erciyes University, Kayseri 38039, TürkiyeGenome and Stem Cell Center (GENKÖK), Erciyes University, Kayseri 38039, Türkiye; Gevher Nesibe Genome and Stem Cell Institute, Erciyes University, Kayseri 38039, TürkiyeGenome and Stem Cell Center (GENKÖK), Erciyes University, Kayseri 38039, Türkiye; Institute of Health Sciences, Erciyes University, Kayseri 38039, TürkiyeGenome and Stem Cell Center (GENKÖK), Erciyes University, Kayseri 38039, Türkiye; Gevher Nesibe Genome and Stem Cell Institute, Erciyes University, Kayseri 38039, TürkiyeDepartment of Molecular Biology and Genetics, Izmir Institute of Technology, Izmir 35430, TürkiyeKANKA Pediatric Bone Marrow Transplantation Center, Department of Pediatric Hematology and Oncology, Faculty of Medicine, Erciyes University, Kayseri 38039, TürkiyeFaculty of Science, Department of Biology, Erciyes University, Kayseri 38039, Türkiye; Genome and Stem Cell Center (GENKÖK), Erciyes University, Kayseri 38039, Türkiye; Corresponding authorSummary: Analysis of the surfaceome of a blood cell subset requires cell sorting, followed by surface protein enrichment. Here, we present a protocol combining magnetically activated cell sorting (MACS) and surface biotinylation of the target cell subset from human peripheral blood mononuclear cells (PBMCs). We describe the steps for isolating target cells and their in-column surface biotinylation, followed by isolation and mass spectrometry analysis of biotinylated proteins. The protocol enables in-column surface biotinylation of specific cell subsets with minimal membrane disruption. : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166722007432Cell BiologyCell IsolationProtein BiochemistryProteomicsMass Spectrometry |
| spellingShingle | Serife Ayaz-Guner Mustafa Burak Acar Dudu Boyvat Huseyin Guner Habibe Bozalan Melis Güzel Selin Kübra Yıldır Nilay Altınsoy Fatma Fındık Musa Karakükçü Servet Özcan Protocol for cell surface biotinylation of magnetic labeled and captured human peripheral blood mononuclear cells STAR Protocols Cell Biology Cell Isolation Protein Biochemistry Proteomics Mass Spectrometry |
| title | Protocol for cell surface biotinylation of magnetic labeled and captured human peripheral blood mononuclear cells |
| title_full | Protocol for cell surface biotinylation of magnetic labeled and captured human peripheral blood mononuclear cells |
| title_fullStr | Protocol for cell surface biotinylation of magnetic labeled and captured human peripheral blood mononuclear cells |
| title_full_unstemmed | Protocol for cell surface biotinylation of magnetic labeled and captured human peripheral blood mononuclear cells |
| title_short | Protocol for cell surface biotinylation of magnetic labeled and captured human peripheral blood mononuclear cells |
| title_sort | protocol for cell surface biotinylation of magnetic labeled and captured human peripheral blood mononuclear cells |
| topic | Cell Biology Cell Isolation Protein Biochemistry Proteomics Mass Spectrometry |
| url | http://www.sciencedirect.com/science/article/pii/S2666166722007432 |
| work_keys_str_mv | AT serifeayazguner protocolforcellsurfacebiotinylationofmagneticlabeledandcapturedhumanperipheralbloodmononuclearcells AT mustafaburakacar protocolforcellsurfacebiotinylationofmagneticlabeledandcapturedhumanperipheralbloodmononuclearcells AT duduboyvat protocolforcellsurfacebiotinylationofmagneticlabeledandcapturedhumanperipheralbloodmononuclearcells AT huseyinguner protocolforcellsurfacebiotinylationofmagneticlabeledandcapturedhumanperipheralbloodmononuclearcells AT habibebozalan protocolforcellsurfacebiotinylationofmagneticlabeledandcapturedhumanperipheralbloodmononuclearcells AT melisguzel protocolforcellsurfacebiotinylationofmagneticlabeledandcapturedhumanperipheralbloodmononuclearcells AT selinkubrayıldır protocolforcellsurfacebiotinylationofmagneticlabeledandcapturedhumanperipheralbloodmononuclearcells AT nilayaltınsoy protocolforcellsurfacebiotinylationofmagneticlabeledandcapturedhumanperipheralbloodmononuclearcells AT fatmafındık protocolforcellsurfacebiotinylationofmagneticlabeledandcapturedhumanperipheralbloodmononuclearcells AT musakarakukcu protocolforcellsurfacebiotinylationofmagneticlabeledandcapturedhumanperipheralbloodmononuclearcells AT servetozcan protocolforcellsurfacebiotinylationofmagneticlabeledandcapturedhumanperipheralbloodmononuclearcells |