Enzymatic amplification and expression of bovine interleukin-1 receptor antagonist cDNA

ELISA for Bovine Interleukin-1 Receptor Antagonist and Its Application to Mastitic Sera and Whey.

“…1A), which coincided with those in previous reports on human IL-1ra [1][2][3][4]. In previous report, we reported that the rbIL1ra inhibited IL-1β activity on A375 cells in a dosedependent manner [9]. In the present study, we confirmed that rbIL-1ra also inhibited IL-1α activity (data not shown), indicating that rbIL-1ra is biologically active.…”

“…We previously reported the sequence of bovine (b) IL-1ra and production of recombinant (r) IL-1ra in E. coli [9]. In the present paper, we describe a sandwich ELISA for bIL1ra using anti-rbIL-1ra rabbit IgG and application of the assay to bovine sera and whey with or without mastitis.…”

“…Production and purification of rbIL-1ra was performed as described previously [9]. Molecular weight of rbIL-1ra was determined by 12.5% sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE) according to the method of Laemmli [10].…”

“…The biological activity of rbIL-1ra was determined by suppression of A375 cell growth inhibition activity of bovine IL-1 [6,11] as described previously [9].…”

Effects of Lipopolysaccharide on Production of Interleukin-1 and Interleukin-6 by Bovine Mammary Epithelial Cells in vitro

“…A 456-bp fragment of the bovine IL-1 receptor antagonist (IL-1ra) and a 552-bp fragment of bovine glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were a m p l i f i e d b y R T -P C R . I L -1 r a ( 5 ' -CACCCCTTGGGAAAGAGACG, 3-CTAGTCCTGCTGG AAGTAGA) [5] and GAPDH (5'-TGGCAAAGTG GACATCGTCG, 3'-GCAGGGATGATATTCTGGGC) oligonucleotides were used as amplification primers. The Cytokines are soluble mediators which have multiple functions and biologic activities.…”