Mice received a single intraperitoneal injection of 30 mg/kg of E. coli LPS (Sigma-Aldrich, St Louis, MO) or PBS as vehicle control and were executed always at ZT4 or 4 hr after the initiation of light into the animal room. At different time points after PBS or LPS administration, BM cells from femurs, tibias, and pelvis were harvested by crushing in PBS containing 2% of FBS and erythrocytes were lysed using a hypotonic buffer from BD Biosciences. Blood was collected by retro-orbital bleeding or cardiac injection. Liver, kidney, and thoracic duct cells were harvested by enzymatic digestion solution with collagenase II (1 mg/mL, Thermo Fisher–Gibco, Waltham, MA) and dispase (5 mg/mL, Gibco, Life Technologies) in a shaking water bath at 37°C for 1 hr. Spleen cells were isolated by scraping with slides in sterile PBS following red blood cell (RBC) lysis (Pharm Lyse; BD Biosciences, San Jose, CA). Extracted LN were derived from the cervical and axillary chains exclusively.
Hypotonic buffer
Manufactured by BD
Hypotonic buffer is a laboratory solution used to dilute and stabilize biological samples. It is designed to create an environment with a lower solute concentration compared to the sample, allowing for controlled osmotic pressure and maintaining the integrity of the sample's cellular structure.
Automatically generated - may contain errors
2 protocols using hypotonic buffer
1
Comprehensive Tissue Immune Profiling After Endotoxin Challenge
2
Systemic Inflammation Induction in Mice
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Mice received a single intraperitoneal injection of 30 mg per Kilo of E. coli LPS (Sigma-Aldrich, St Louis, MO) or PBS as vehicle control and were executed always at ZT4 or 4 hours after the initiation of light into the animal room. At different time points after PBS or LPS administration, BM cells from femurs, tibias and pelvis were harvested by crushing in PBS containing 2% of FBS and erythrocytes were lysed using a hypotonic buffer from BD Biosciences. Blood was collected by retro-orbital bleeding or cardiac injection. Liver, kidney and thoracic duct cells were harvested by enzymatic digestion solution with collagenase II (1 mg/mL, ThermoFisher Gibco, Waltham, MA) and dispase (5 mg/mL, Gibco, Life Technologies) in a shaking water bath at 37 0 C for 1h. Spleen cells were isolated by scraping with slides in sterile PBS following red blood cell (RBC) lysis (Pharm Lyse TM ; BD Bioscience, San Jose, CA).
Extracted LN were derived from the cervical and axillary chains exclusively.
Extracted LN were derived from the cervical and axillary chains exclusively.
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