An Improved m6A-ELISA for Quantifying N6-methyladenosine in Poly(A)-purified mRNAs
N6-methyladenosine (m6A) is an abundant internal mRNA modification with roles in regulating cellular and organismal physiology, including development, differentiation, and disease. The deposition of m6A is highly regulated, with various m6A levels across different environmental conditions, cellular...
Saved in:
| Main Authors: | , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Bio-protocol LLC
2025-06-01
|
| Series: | Bio-Protocol |
| Online Access: | https://bio-protocol.org/en/bpdetail?id=5359&type=0 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1839655628339937280 |
|---|---|
| author | Wei Chan Waleed Albihlal Folkert Van Werven |
| author_facet | Wei Chan Waleed Albihlal Folkert Van Werven |
| author_sort | Wei Chan |
| collection | DOAJ |
| description | N6-methyladenosine (m6A) is an abundant internal mRNA modification with roles in regulating cellular and organismal physiology, including development, differentiation, and disease. The deposition of m6A is highly regulated, with various m6A levels across different environmental conditions, cellular states, and cell types. Available methods for measuring bulk m6A levels are often time-consuming, have low throughput, and/or require specialized instrumentation or data analyses. Here, we present a detailed protocol for measuring bulk m6A levels in purified poly(A) RNA samples with m6A-ELISA using a standard-based approach. Critical steps of the protocol are highlighted and optimized, including poly(A) RNA quality controls and antibody specificity testing. The protocol is fast, scalable, adaptable, and cost-effective. It does not require specialized instrumentation, training, or skills in data analysis. We have successfully tested this protocol on mRNAs isolated from budding yeast and mouse cell lines. |
| format | Article |
| id | doaj-art-bd176a38d19f45e19f74fc38d85d66d7 |
| institution | Matheson Library |
| issn | 2331-8325 |
| language | English |
| publishDate | 2025-06-01 |
| publisher | Bio-protocol LLC |
| record_format | Article |
| series | Bio-Protocol |
| spelling | doaj-art-bd176a38d19f45e19f74fc38d85d66d72025-06-25T06:53:47ZengBio-protocol LLCBio-Protocol2331-83252025-06-01151210.21769/BioProtoc.5359An Improved m6A-ELISA for Quantifying N6-methyladenosine in Poly(A)-purified mRNAsWei Chan0Waleed Albihlal1Folkert Van Werven2Cell Fate and Gene Regulation Laboratory, The Francis Crick Institute, London, UKCell Fate and Gene Regulation Laboratory, The Francis Crick Institute, London, UKCell Fate and Gene Regulation Laboratory, The Francis Crick Institute, London, UKN6-methyladenosine (m6A) is an abundant internal mRNA modification with roles in regulating cellular and organismal physiology, including development, differentiation, and disease. The deposition of m6A is highly regulated, with various m6A levels across different environmental conditions, cellular states, and cell types. Available methods for measuring bulk m6A levels are often time-consuming, have low throughput, and/or require specialized instrumentation or data analyses. Here, we present a detailed protocol for measuring bulk m6A levels in purified poly(A) RNA samples with m6A-ELISA using a standard-based approach. Critical steps of the protocol are highlighted and optimized, including poly(A) RNA quality controls and antibody specificity testing. The protocol is fast, scalable, adaptable, and cost-effective. It does not require specialized instrumentation, training, or skills in data analysis. We have successfully tested this protocol on mRNAs isolated from budding yeast and mouse cell lines.https://bio-protocol.org/en/bpdetail?id=5359&type=0 |
| spellingShingle | Wei Chan Waleed Albihlal Folkert Van Werven An Improved m6A-ELISA for Quantifying N6-methyladenosine in Poly(A)-purified mRNAs Bio-Protocol |
| title | An Improved m6A-ELISA for Quantifying N6-methyladenosine in Poly(A)-purified mRNAs |
| title_full | An Improved m6A-ELISA for Quantifying N6-methyladenosine in Poly(A)-purified mRNAs |
| title_fullStr | An Improved m6A-ELISA for Quantifying N6-methyladenosine in Poly(A)-purified mRNAs |
| title_full_unstemmed | An Improved m6A-ELISA for Quantifying N6-methyladenosine in Poly(A)-purified mRNAs |
| title_short | An Improved m6A-ELISA for Quantifying N6-methyladenosine in Poly(A)-purified mRNAs |
| title_sort | improved m6a elisa for quantifying n6 methyladenosine in poly a purified mrnas |
| url | https://bio-protocol.org/en/bpdetail?id=5359&type=0 |
| work_keys_str_mv | AT weichan animprovedm6aelisaforquantifyingn6methyladenosineinpolyapurifiedmrnas AT waleedalbihlal animprovedm6aelisaforquantifyingn6methyladenosineinpolyapurifiedmrnas AT folkertvanwerven animprovedm6aelisaforquantifyingn6methyladenosineinpolyapurifiedmrnas AT weichan improvedm6aelisaforquantifyingn6methyladenosineinpolyapurifiedmrnas AT waleedalbihlal improvedm6aelisaforquantifyingn6methyladenosineinpolyapurifiedmrnas AT folkertvanwerven improvedm6aelisaforquantifyingn6methyladenosineinpolyapurifiedmrnas |