Sdf 1 alpha cxcl12a human elisa kit

Analysis was performed on a subset of the full cohort. Biopsy samples were homogenized using TissueLyzer (21 Hz, 1.5 min) in lysis buffer (Bio-Plex cell lysis kit, #171304011, according to the supplier’s instruction). Samples were frozen at −80°C for 2 hours and then thawed and centrifugated (4 min, 4500g) at 4°C. Supernatants were collected and protein concentration was measured using a bicinchoninic (BCA) assay. Samples were diluted to 3 μg/μl and stored at −80°C until further use. Cytokines were measured using a custom Bio-Plex Multiplex Immunoassay System (Bio-Rad) on samples diluted 1:4 in water according to the manufacturer’s instructions. CXCL12 was measured in skeletal muscle lysate diluted 1:5 using SDF-1 alpha/CXCL12A human ELISA kit (Invitrogen, EHCXCL12A) and SDF-1 beta/CXCL12B human ELISA kit (Invitrogen EHCXCL12B). Values below detection were imputed with the minimum concentration measurable by the kit according to the manufacturer.

Heparin plasma collected during the exercise day was used to investigate cytokine levels. Analysis was performed on a subset of the full cohort. IL-6, creatine, and CXCL2 levels were investigated using the following kits: human IL-6 ELISA MAX deluxe (BioLegend, #430505), human CKM ELISA kit (Abcam, ab185988), and human MIP2 ELISA kit (CXCL2) (Abcam, ab184862), according to the manufacturer’s instructions. A human acute phase 5-plex panel (Bio-Rad, #171a4008m, lot 5036639) was used to measure ferritin, fibrinogen, procalcitonin, serum amyloid A, and tissue plasminogen activator, with a sample dilution of 1:100. A human acute phase 4-plex panel (Bio-Rad, #171a4009m) was used to measure α-2-macroglobin, CRP, haptoglobin, and serum amyloid P, with a sample dilution of 1:10,000. CXCL12 was measured in plasma diluted 1:2 using SDF-1 alpha/CXCL12A human ELISA kit (Invitrogen, lot EHCXCL12A) and SDF-1 beta/CXCL12B human ELISA kit (Invitrogen, lot EHCXCL12B). All assays were performed according to the manufacturer’s instructions. Values below detection were imputed with the minimum concentration measurable by the kit according to the manufacturer. Blood measurements were tested for outliers using interquartile range and analyzed using two-way repeated analysis of variance (ANOVA) in R v4.0.4. P < 0.05 was considered significant.