biotinylated anti als  (R&D Systems)

Journal: Toxicological Sciences

Article Title: Suppression of Insulin-Like Growth Factor Acid-Labile Subunit Expression--A Novel Mechanism for Deoxynivalenol-Induced Growth Retardation

doi: 10.1093/toxsci/kfp225

Figure Lengend Snippet: DON consumption reduces hepatic IGFALS mRNA in mice. Mice were fed diets as described in Figure 1 and groups sacrificed at intervals. Total mRNA from liver was analyzed by real-time PCR. Data are mean ± SEM (n = 6–8) of fold changes relative to vehicle control arbitrarily set at 100. Means with asterisks differ from control mean at specified time interval (p < 0.05).

Article Snippet: Plates were washed and wells incubated with 100 μl of 200 ng/ml IGFALS biotinylated antibody (BAF 1436; R & D systems) dissolved in PBS containing 2% (vol/vol) goat serum (Sigma) and 0.5% (vol/vol) Tween-20, for 2 h at room temperature.

Techniques: Real-time Polymerase Chain Reaction, Control

Journal: Toxicological Sciences

Article Title: Suppression of Insulin-Like Growth Factor Acid-Labile Subunit Expression--A Novel Mechanism for Deoxynivalenol-Induced Growth Retardation

doi: 10.1093/toxsci/kfp225

Figure Lengend Snippet: DON consumption reduces circulating IGF1 and IGFALS in mice. Mice were fed diets as described in Figure 1. Plasma was analyzed for (A) IGF1 and (B) IGFALS at intervals by ELISA. Data are mean ± SEM. (n = 6–8). Means with asterisks differ from control mean at the designated time interval (p < 0.05).

Article Snippet: Plates were washed and wells incubated with 100 μl of 200 ng/ml IGFALS biotinylated antibody (BAF 1436; R & D systems) dissolved in PBS containing 2% (vol/vol) goat serum (Sigma) and 0.5% (vol/vol) Tween-20, for 2 h at room temperature.

Techniques: Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Control

Journal: Toxicological Sciences

Article Title: Suppression of Insulin-Like Growth Factor Acid-Labile Subunit Expression--A Novel Mechanism for Deoxynivalenol-Induced Growth Retardation

doi: 10.1093/toxsci/kfp225

Figure Lengend Snippet: Dose-dependent suppression of hepatic IGFALS mRNA in mice following acute oral exposure to DON. Mice were orally gavaged with various doses of DON (0 PBS, 0.1–12.5 mg/kg bw) and liver sections collected 2 h later. Total mRNA was isolated and analyzed by real-time PCR. Data are mean ± SEM (n = 5) of mRNA fold change relative to a vehicle treatment designated as 100. Means with asterisk differ from vehicle (p < 0.05).

Article Snippet: Plates were washed and wells incubated with 100 μl of 200 ng/ml IGFALS biotinylated antibody (BAF 1436; R & D systems) dissolved in PBS containing 2% (vol/vol) goat serum (Sigma) and 0.5% (vol/vol) Tween-20, for 2 h at room temperature.

Techniques: Isolation, Real-time Polymerase Chain Reaction

Journal: Toxicological Sciences

Article Title: Suppression of Insulin-Like Growth Factor Acid-Labile Subunit Expression--A Novel Mechanism for Deoxynivalenol-Induced Growth Retardation

doi: 10.1093/toxsci/kfp225

Figure Lengend Snippet: Acute oral DON exposure differentially affects hepatic mRNA expression of IGF1 ternary complex partners. Mice were orally gavaged with 12.5 mg/kg bw DON or PBS vehicle treatment (VH) and then treated with GH i.p 2 h later. At 3 and 6 h after DON administration, liver was analyzed by real-time PCR for (A) IGFALS, (B) IGF1, and (C) IGFBP3. Data are mean ± SEM (n = 4) of mRNA change in target relative to VH. Means with asterisks differ from VH at the specified time interval (p < 0.05).

Article Snippet: Plates were washed and wells incubated with 100 μl of 200 ng/ml IGFALS biotinylated antibody (BAF 1436; R & D systems) dissolved in PBS containing 2% (vol/vol) goat serum (Sigma) and 0.5% (vol/vol) Tween-20, for 2 h at room temperature.

Techniques: Expressing, Real-time Polymerase Chain Reaction

Journal: Toxicological Sciences

Article Title: Suppression of Insulin-Like Growth Factor Acid-Labile Subunit Expression--A Novel Mechanism for Deoxynivalenol-Induced Growth Retardation

doi: 10.1093/toxsci/kfp225

Figure Lengend Snippet: Acute oral DON exposure suppresses IGFALS mRNA expression in livers of control- and GH-treated mice. Mice were orally gavaged with DON 12.5 mg/kg bw (DON) or PBS (VH) and later treated twice with GH or NaHCO3 buffer (CON) i.p, at 0.25 and 2 h after DON exposure. Mice were sacrificed 4 h after DON administration and liver sections collected and analyzed for IGFALS mRNA expression by real-time PCR. Data are mean ± SEM (n = 4) of mRNA fold change relative to VH/CON group arbitrarily designated as 100. Means without the same letter differ (p < 0.05).

Article Snippet: Plates were washed and wells incubated with 100 μl of 200 ng/ml IGFALS biotinylated antibody (BAF 1436; R & D systems) dissolved in PBS containing 2% (vol/vol) goat serum (Sigma) and 0.5% (vol/vol) Tween-20, for 2 h at room temperature.

Techniques: Expressing, Control, Real-time Polymerase Chain Reaction

Journal: Toxicological Sciences

Article Title: Suppression of Insulin-Like Growth Factor Acid-Labile Subunit Expression--A Novel Mechanism for Deoxynivalenol-Induced Growth Retardation

doi: 10.1093/toxsci/kfp225

Figure Lengend Snippet: Proposed mechanism for DON-induced growth impairment. DON induces systemic upregulation of several proinflammatory cytokines capable of inducing SOCS expression. SOCS can interfere with growth hormone receptor (GHR) signaling by (1) impairing GH-induced STAT or JAK phosphorylation of GHR or (2) mediating proteasomal degradation of GHR. A potential outcome of these events is reduction in GH-induced hepatic IGFALS mRNA expression and ultimately circulating IGFALS. The loss of IGFALS results in increased IGF1 degradation and the reduced circulating levels of this hormone ultimately causing growth retardation.

Article Snippet: Plates were washed and wells incubated with 100 μl of 200 ng/ml IGFALS biotinylated antibody (BAF 1436; R & D systems) dissolved in PBS containing 2% (vol/vol) goat serum (Sigma) and 0.5% (vol/vol) Tween-20, for 2 h at room temperature.

Techniques: Expressing, Phospho-proteomics