Version 1
: Received: 5 November 2024 / Approved: 6 November 2024 / Online: 6 November 2024 (17:07:06 CET)
How to cite:
Negreros, M.; Hernández, F.; Mendoza-Milla, C.; Cisneros, J.; Morales-Hurtado, F.; Flores-Suárez, L. F. Time-Course and Fraction Analysis in Western Blot Reveals Improved Approaches for Evaluating Neutrophil Extracellular Trap-Associated Proteins. Preprints2024, 2024110426. https://doi.org/10.20944/preprints202411.0426.v1
Negreros, M.; Hernández, F.; Mendoza-Milla, C.; Cisneros, J.; Morales-Hurtado, F.; Flores-Suárez, L. F. Time-Course and Fraction Analysis in Western Blot Reveals Improved Approaches for Evaluating Neutrophil Extracellular Trap-Associated Proteins. Preprints 2024, 2024110426. https://doi.org/10.20944/preprints202411.0426.v1
Negreros, M.; Hernández, F.; Mendoza-Milla, C.; Cisneros, J.; Morales-Hurtado, F.; Flores-Suárez, L. F. Time-Course and Fraction Analysis in Western Blot Reveals Improved Approaches for Evaluating Neutrophil Extracellular Trap-Associated Proteins. Preprints2024, 2024110426. https://doi.org/10.20944/preprints202411.0426.v1
APA Style
Negreros, M., Hernández, F., Mendoza-Milla, C., Cisneros, J., Morales-Hurtado, F., & Flores-Suárez, L. F. (2024). Time-Course and Fraction Analysis in Western Blot Reveals Improved Approaches for Evaluating Neutrophil Extracellular Trap-Associated Proteins. Preprints. https://doi.org/10.20944/preprints202411.0426.v1
Chicago/Turabian Style
Negreros, M., Fernando Morales-Hurtado and Luis Felipe Flores-Suárez. 2024 "Time-Course and Fraction Analysis in Western Blot Reveals Improved Approaches for Evaluating Neutrophil Extracellular Trap-Associated Proteins" Preprints. https://doi.org/10.20944/preprints202411.0426.v1
Abstract
Neutrophil extracellular traps (NETs) dysregulation is implicated in various diseases, highlighting the need for accurate quantification methods. These methods must account for the unique nature of NETs, where nuclear DNA and intracellular proteins translocate to the extracellular space. This study addresses technical approaches in the Western blot quantification of NETs-associated proteins (NAPs). Neutrophils were isolated from healthy donors, stimulated with phorbol-myristate acetate (PMA), and the expression of cathelicidin antimicrobial protein LL-37 (LL-37/hCAP18), glyceraldehyde 3-phosphate dehydrogenase (GAPDH), and β-actin was analyzed over time. LL-37/hCAP18 exhibited temporal and fraction-specific distribution patterns, while the evaluation of GAPDH and β-actin as loading controls emphasized the need for more precise tailoring when studying NETs. This work aims to improve understanding of the techniques used to measure them to make more accurate comparisons in the area of NETs research. Western blot emerges as a robust and accessible tool, shedding light on the dynamics of NAPs.
Keywords
Neutrophil Extracellular Traps; Western Blot; LL-37; GAPDH; b-actin; Loading Controls
Subject
Biology and Life Sciences, Immunology and Microbiology
Copyright:
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
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