Amphipod protein content under simulated invasion stress in wetlands

DOI

Every two weeks during the 6-week acclimation period, 8 adults were randomly selected from each tank for protein analysis (n = 8). Individuals were sacrificed in dry ice and stored at -80 °C. Whole frozen amphipods were prepared for protein quantification using a colorimetric assay (Thermo Scientific Pierce BCA Protein Assay Kit). Samples were homogenized with a Pro200 Bio-Gen Series homogenizer for 10 seconds on ice with the homogenization buffer (100 mmol l⁻¹ Tris-HCl, pH 7.5; 0.1% SDS (w/v), 0.5 mol l⁻¹ EDTA) containing a combination of protease inhibitors: 0.7 μg ml⁻¹ pepstatin A, 0.5 μg ml⁻¹ leupeptin, 1 μg ml⁻¹ aprotinin and 20 μg ml⁻¹ phenylmethylsulfonyl fluoride at a dilution ratio of 1:20. In several instances, protein concentration at a 1:20 dilution was too high for the range of measurement: seven individuals were further diluted to 1:30 and two individuals were further diluted to 1:35. Homogenates were centrifuged at 13,000 g for 10 minutes. 50 μl of the resulting supernatant was transferred into a new 0.5 ml tube to use for total protein quantification. 10 μl of each sample was added to triplicate wells in 96 well plates. 200 μl of the BCA reagent was added to the wells and then placed on the shaker for 30 seconds. Plates were heated at 37 °C for 30 minutes and placed on the spectrometer at 562 nm. Protein values were standardized with 2 mg ml⁻¹ BSA standard.

Funding:* Community function under Phragmites invasion in Suisun Marsh (https://www.seacrlab.com/), Award: Delta Stewardship Council DSF R/SF-107

Identifier
DOI https://doi.org/10.1594/PANGAEA.973194
Related Identifier IsPartOf https://doi.org/10.1594/PANGAEA.973173
Metadata Access https://ws.pangaea.de/oai/provider?verb=GetRecord&metadataPrefix=datacite4&identifier=oai:pangaea.de:doi:10.1594/PANGAEA.973194
Provenance
Creator Tanner, Richelle L ORCID logo; Haworth, Lorna; Nancollas, Sarah; Landa, Susie; Todgham, Anne E
Publisher PANGAEA
Publication Year 2024
Rights Creative Commons Attribution 4.0 International; https://creativecommons.org/licenses/by/4.0/
OpenAccess true
Representation
Resource Type Dataset
Format text/tab-separated-values
Size 3393 data points
Discipline Biospheric Sciences; Ecology; Geosciences; Life Sciences; Medicine; Medicine and Health; Natural Sciences; Physiology
Spatial Coverage (-121.983 LON, 38.118 LAT); Suisun Marsh, California, United States