Author: Jacob E. Choby; Hanna B. Buechi; Allison J. Farrand; Eric P. Skaar; Matthew F. Barber
Title: Molecular basis for the evolution of species-specific hemoglobin capture by pathogenic Staphylococcus Document date: 2018_6_7
ID: ieh5cvw1_51
Snippet: The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. . https://doi.org/10.1101/339705 doi: bioRxiv preprint by densitometric analysis with Image J (NIH). Because of variation in stain intensity across gels, all 408 comparisons were made within the same gel, and relative density was calculated for each biological 409 replicate within the same gel; the comparison was either to human hemoglobin or wildtype IsdB,.....
Document: The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. . https://doi.org/10.1101/339705 doi: bioRxiv preprint by densitometric analysis with Image J (NIH). Because of variation in stain intensity across gels, all 408 comparisons were made within the same gel, and relative density was calculated for each biological 409 replicate within the same gel; the comparison was either to human hemoglobin or wildtype IsdB, 410 depending on assay. Additionally, PBS only samples and S. aureus ΔisdB::erm were used to verify that 411 hemoglobin binding in this assay is IsdB dependent as previously observed [Supplemental Figure 1 ] (11, 412 13 MnCl 2 , 100 μM CaCl 2 , and 1 mM MgCl 2 (all from Fisher) to restore non-iron cations, 1.5 mM EDDHA to 422 chelate any remaining free iron, and 2.5 µg/ml of recombinant purified hemoglobin as the sole iron source. 423
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