The objective was to provide an updated profile of the bovine acute-phase response to include recent advancements in technologies and expanded hematological, cytokine, and serum chemistry variables. Beef steers (n = 32; body weight [BW] = 251 ± 19.5 kg) were fitted with indwelling jugular catheters 1 d before lipopolysaccharide (LPS; 0.25 µg LPS/kg BW from Escherichia coli O111:B4) administration to facilitate serial blood collection. Rectal temperature was measured using indwelling probes, and ocular temperature was measured using infrared thermal imaging. Blood samples were collected for subsequent analysis of serum chemistry, hematology, and cytokine concentrations. Pearson correlation of rectal temperature and ocular infrared temperature was 0.61 (P < 0.01) and the Spearman correlation coefficient was 0.56 (P < 0.01). Interactions of hour × method were observed for ocular and rectal measurements of body temperature in response to endotoxin exposure. Maximum observed temperature was 39.6 °C at 2.5 h for both rectal and ocular measurements. Body temperature differed by method at hours 0.5, 2.5, 4.5, 7.5, 12.5, 36.5, and 47.5 (P < 0.01), but were not different otherwise. All variables of serum chemistry and complete blood count were influenced by LPS administration, except creatinine, serum glucose, and percent basophils (P ≤ 0.02). Alanine aminotransferase and alkaline phosphatase peaked at hour 2 relative to LPS administration, returned to baseline at hour 12 and continued to decrease below the baseline value at hour 48 (P < 0.01). Total protein concentration decreased 3% in response to LPS (P = 0.01). Total white blood cell count decreased 75% after LPS administration at hour 1 (P < 0.01). Lymphocyte count recovered to baseline at hour 6; sooner than neutrophil count at hour 36. Serum cortisol concentration increased 294% relative to baseline at hour 1 followed by a sustained decrease and return to normal concentration at hour 4 (P < 0.01). Additionally, circulating cytokine concentrations changed with time in response to the LPS challenge, excluding aFGF, bFGF, IGF-1, IL-2, IL-4, MCP-1, and ANG-1 (P ≤ 0.08). Maximum observed concentration of TNF-α at hour 1 was 117% greater than the pre-challenge value (P < 0.01). Data presented herein add to existing works to understand the endocrine and immune responses of beef steers administered exogenous LPS, and incorporate recent technologies, additional biomarkers, and an expanded cytokine profile that can be used as referential data in future research.