lif recombinant human protein (hlif  (Thermo Fisher)

Journal: Communications Biology

Article Title: An engineered ligand trap inhibits leukemia inhibitory factor as pancreatic cancer treatment strategy

doi: 10.1038/s42003-021-01928-2

Figure Lengend Snippet: a Yeast-displayed eLIFR containing the CBM I–Ig-like–CBM II domains (blue circles) has a slightly higher affinity for LIF-His than eLIFR CBM I–Ig-like domains (gray squares). Data are the mean ± SD of triplicate measurements. Data from individual experiments are shown as faint symbols. b Schematic of hLIFR-Fc and eLIFR-Fc. The three N-terminal domains, CBM I–Ig-like–CBM II, are fused to an hIgG1 Fc-domain, with the engineered Ig-like domain shown in dark teal for eLIFR-Fc. c Versus hLIFR-Fc, eLIFR-Fc remains more strongly bound to yeast-displayed LIF after overnight incubation with soluble LIF competitor, indicating a slower off-rate. * P = 0.024, ** P = 0.0024 versus the corresponding hLIFR-Fc condition, two-tailed unpaired Student’s t test. Data are mean ± SD ( n = 3). d KinExA data showing that recombinant eLIFR-Fc (blue triangles) has higher affinity to soluble hLIF-His than hLIFR-Fc (brown circles). Data are the mean of duplicate measurements. Data from individual experiments are shown as faint symbols. e hLIFR-Fc, eLIFR-Fc (bivalent and one-arm), and D25 antibody compete LIF away from WT LIFR. ns not significant, P = 0.03 for one-arm eLIFR-Fc and P = 0.04 for eLIFR-Fc versus hLIFR-Fc, two-tailed unpaired Student’s t test. f Both hLIFR-Fc and eLIFR-Fc (bivalent and one-arm) compete LIF away from WT gp130, but the D25 mAb does not and appears to increase binding, perhaps due to complex stabilization or more avid LIF binding. P = 0.02, ** P = 0.006, or *** P < = 0.0002 versus hLIFR-Fc or eLIFR-Fc (as indicated), two-tailed unpaired Student’s t test. For e and f , data are mean ± SD ( n ≥ 3 independent experiments).

Article Snippet: Recombinant hLIF-His (14890-H08H-20, Sino Biological Inc.), hLIF (untagged) (14890-HNAH-50, Sino Biological Inc.), mLIF-His (ABIN2215872, Antibodies-Online), hOSM-His (10425-H08H-20, Sino Biological Inc.), and hCTF-1-Fc (16013-H01H-20, Sino Biological Inc.) were purchased for use.

Techniques: Incubation, Two Tailed Test, Recombinant, Binding Assay

Journal: The Journal of clinical endocrinology and metabolism

Article Title: Measurement of leukemia inhibitory factor in biological fluids by radioimmunoassay.

doi: 10.1210/jcem.83.4.4702

Figure Lengend Snippet: FIG. 1. Cross-reaction in mLIF and hLIF RIAs. Serial dilutions of a mixture of IL-1, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12, G-CSF, IFN, and TGF at the highest dose (131 ng/mL; R&D Systems); hOSM at the highest dose 100 ng/mL; and mLIF at the highest dose of 100 ng/mL were applied to the hLIF RIA (top). Serial dilutions of hLIF (highest dose, 100 ng/mL), mIL-6 (highest dose, 100 ng/mL), and hOSM (high- est dose, 100 ng/mL) were applied to the mLIF RIA (bottom).

Article Snippet: Materials and Methods Radioiodination of LIF Escherichia coli-derived recombinant hLIF, mLIF, goat polyclonal antihLIF antibody, and anti-mLIF antibody were commercially purchased (R&D Systems, Minneapolis, MN).

Techniques:

Journal: The Journal of clinical endocrinology and metabolism

Article Title: Measurement of leukemia inhibitory factor in biological fluids by radioimmunoassay.

doi: 10.1210/jcem.83.4.4702

Figure Lengend Snippet: FIG. 2. Size-exclusion profile of [125I]LIF in serum. The Ultrogel AcA 22 (1.5 3 95-cm) column was equilibrated and run in 0.01 mol/L PBS at 1.5 mL/fraction. Fraction number: void volume, 33; total volume, 105; and 31 kDa, 72. A, [125I]hLIF in the presence or absence of human nonpregnant serum. B, [125I]hLIF in human pregnancy serum with or without a 40-fold excess of unlabeled hLIF. C, [125I]mLIF incubated in pregnant mouse serum.

Article Snippet: Materials and Methods Radioiodination of LIF Escherichia coli-derived recombinant hLIF, mLIF, goat polyclonal antihLIF antibody, and anti-mLIF antibody were commercially purchased (R&D Systems, Minneapolis, MN).

Techniques: Incubation