goat anti mouse igf 1  (R&D Systems)

Journal: PLoS ONE

Article Title: E-Peptides Control Bioavailability of IGF-1

doi: 10.1371/journal.pone.0051152

Figure Lengend Snippet: Exons 1 and 2 are transcribed from different promoters. Differential splicing gives rise to two different signal peptides (SP1 and SP2), which include a common C-terminal sequence encoded by Exon 3. Exon 3 also encodes the N-terminal part of the mature IGF-1 B chain. Exon 4 encodes the remaining mature IGF-1 protein (B,C,A and D chains), and also encodes the common N-terminal sequence of the E-peptides. Differential splicing excluding Exon 5 gives rise to the IGF-1Ea propeptide, or a longer IGF-1Eb propeptide when Exon 5 is included. Protease cleavage (arrowheads) removes the E peptides to produce the mature IGF-1 protein.

Article Snippet: In the case of IGF-1, the primary antibody was a goat anti-IGF-1 antibody (Sigma, I-8773) was used at a 1∶1000 dilution and the secondary antibody, donkey anti-goat IgG-HRP (Santa Cruz, sc-2020) was used at 1∶10.000.

Techniques: Sequencing

Journal: PLoS ONE

Article Title: E-Peptides Control Bioavailability of IGF-1

doi: 10.1371/journal.pone.0051152

Figure Lengend Snippet: A) Western blot analysis of IGF-1 transgene levels in quadriceps muscle of 3 months old male mice. B) Total IGF-1 levels in the blood serum of 3 months old transgenic male mice compared to WT littermates as determined by ELISA.

Article Snippet: In the case of IGF-1, the primary antibody was a goat anti-IGF-1 antibody (Sigma, I-8773) was used at a 1∶1000 dilution and the secondary antibody, donkey anti-goat IgG-HRP (Santa Cruz, sc-2020) was used at 1∶10.000.

Techniques: Western Blot, Transgenic Assay, Enzyme-linked Immunosorbent Assay

Journal: PLoS ONE

Article Title: E-Peptides Control Bioavailability of IGF-1

doi: 10.1371/journal.pone.0051152

Figure Lengend Snippet: A) Growth medium (10 uL) from transiently transfected HEK 293 cells (IGF-1 levels normalised to 200 ng/mL). B) Binding of IGF-1 propeptides to positively (amine) (lanes 2–4) and negatively (carboxyl) (lanes 6–8) charged tissue culture plates. The control lane (9) is a mixture of growth media from IGF-1-stop and IGF-1EbCD transfected cells.

Article Snippet: In the case of IGF-1, the primary antibody was a goat anti-IGF-1 antibody (Sigma, I-8773) was used at a 1∶1000 dilution and the secondary antibody, donkey anti-goat IgG-HRP (Santa Cruz, sc-2020) was used at 1∶10.000.

Techniques: Transfection, Binding Assay

Journal: PLoS ONE

Article Title: E-Peptides Control Bioavailability of IGF-1

doi: 10.1371/journal.pone.0051152

Figure Lengend Snippet: Binding of IGF-1 isoforms to heparin coated agarose beads (lanes 2–4) and control agarose beads (lanes 6–8). The control lane (9) is the growth medium from IGF-1EbCD transfected cells.

Article Snippet: In the case of IGF-1, the primary antibody was a goat anti-IGF-1 antibody (Sigma, I-8773) was used at a 1∶1000 dilution and the secondary antibody, donkey anti-goat IgG-HRP (Santa Cruz, sc-2020) was used at 1∶10.000.

Techniques: Binding Assay, Transfection

Journal: PLoS ONE

Article Title: E-Peptides Control Bioavailability of IGF-1

doi: 10.1371/journal.pone.0051152

Figure Lengend Snippet: Western blot analysis of IGF-1 binding. Lanes 1–4: growth media from HEK 293 cells transfected with IGF-1 expression plasmids encoding either the mature peptide (lane 2) or cleavage deficient IGF-1 propeptides (lanes 3 and 4). Lanes 5–8: same growth media after incubation with decellularizsed tissue. Lanes 9–12: IGF-1 binding to decellularized lung tissue. Lanes 13–16: IGF-1 binding to decellularizsed skeletal muscle tissue.

Article Snippet: In the case of IGF-1, the primary antibody was a goat anti-IGF-1 antibody (Sigma, I-8773) was used at a 1∶1000 dilution and the secondary antibody, donkey anti-goat IgG-HRP (Santa Cruz, sc-2020) was used at 1∶10.000.

Techniques: Western Blot, Binding Assay, Transfection, Expressing, Incubation

Journal: PLoS ONE

Article Title: E-Peptides Control Bioavailability of IGF-1

doi: 10.1371/journal.pone.0051152

Figure Lengend Snippet: A–D) Decellularized lung tissue was sectioned and incubated with growth media from HEK 293 cells transfected with IGF-1 expression plasmids (see for details). Bound IGF-1 was visualized by immunostaining using anti-IGF-1 antibody. E) Quantification of the number of IGF-1 loci. Data is presented as mean (SE) for 20 biological replicates. Two stars corresponds to P<0.01, three stars correspond to P<0.001.

Article Snippet: In the case of IGF-1, the primary antibody was a goat anti-IGF-1 antibody (Sigma, I-8773) was used at a 1∶1000 dilution and the secondary antibody, donkey anti-goat IgG-HRP (Santa Cruz, sc-2020) was used at 1∶10.000.

Techniques: Incubation, Transfection, Expressing, Immunostaining