irrelevant control antibody 13c4  (GenScript corporation)

Journal: Journal of Lipid Research

Article Title: Prevention of renal apoB retention is protective against diabetic nephropathy: role of TGF-β inhibition [S]

doi: 10.1194/jlr.M078204

Figure Lengend Snippet: Effect of TGF-β inhibition. Biglycan wild-type (Bgn+/+; solid symbols) and biglycan-deficient (Bgn−/−; open symbols) male Ldlr−/− mice were injected with STZ and concurrently injected with TGF-β-neutralizing antibody, 1D11 (triangles), or irrelevant control antibody, 13C4 (squares), at 2 mg/kg ip. One week later, all mice were placed on a 0.12% cholesterol diet. A: Blood glucose was measured at the indicated times (mean ± SEM is shown for N = 13–26 per group). There were no significant differences between groups. B: Total TGF-β was measured at least weekly. The 1D11 or 13C4 were reinjected every 8 weeks as indicated by arrows. Data shown is mean ± SEM for N = 17–27 per group; not every mouse was bled at each time point so the number of mice at each time point varies from 2 to 27 per group. P < 0.001 for 13C4-treated biglycan wild-type versus 13C4-treated biglycan-deficient mice; P < 0.001 for 1D11 versus 13C4 treatment within each genotype. The curves for 1D11-treated mice overlap. C: Mice were placed individually in metabolic cages every 4 weeks for the collection of 24 h urine samples for the determination of UAE/urinary creatinine excretion. Mean ± SEM is shown for N = 6–10 per group at each time point. *P < 0.001 for effect of antibody. D: Renal sections were stained with PAS and scored using a semi-quantitative scale by two observers blinded to group with scores averaged. Data shown is the mean ± SEM for N = 7–10 per group. Groups without a common letter are significantly different.

Article Snippet: In some experiments, mice were concurrently injected intraperitoneally with 2 mg/kg body weight TGF-β-neutralizing antibody, 1D11 (R&D Systems, Minneapolis, MN), or irrelevant control antibody, 13C4 (Genscript, Piscataway, NJ), on the first day of STZ injections.

Techniques: Inhibition, Injection, Staining

Journal: Journal of Lipid Research

Article Title: Prevention of renal apoB retention is protective against diabetic nephropathy: role of TGF-β inhibition [S]

doi: 10.1194/jlr.M078204

Figure Lengend Snippet: Renal lipid accumulation. Biglycan wild-type (Bgn+/+) and biglycan-deficient (Bgn−/−) male Ldlr−/− mice were injected with STZ and concurrently injected with TGF-β-neutralizing antibody, 1D11 (solid bars), or irrelevant control antibody, 13C4 (open bars), at 2 mg/kg ip. One week later, all mice were placed on a 0.12% cholesterol diet. A: Total renal protein was immunoblotted for apoB; actin is shown as the loading control. Shown are two mice per group representative of N = 6 per group. Molecular mass markers are shown to the left (in kDa). B: Blots were analyzed by densitometry. Mean ± SEM is shown. *P = 0.002 for antibody effect. C: Mesangial cells isolated from biglycan-deficient or biglycan wild-type mice were treated with TGF-β (2 ng/ml) or vehicle and with 1D11 or 13C4 (10 μg/ml) for 24 h and then washed and incubated for 4 h with Alexa Fluor-labeled LDL; shown is Alexa Fluor intensity normalized to DAPI area. Mean ± SEM is shown for N = 5. *P ≤ 0.05 or **P < 0.0001 for antibody effect; aP < 0.05 for genotype effect. D: Cell protein from parallel wells was immunoblotted for biglycan; actin is shown as the loading control. The blot shown is representative of three separate experiments.

Article Snippet: In some experiments, mice were concurrently injected intraperitoneally with 2 mg/kg body weight TGF-β-neutralizing antibody, 1D11 (R&D Systems, Minneapolis, MN), or irrelevant control antibody, 13C4 (Genscript, Piscataway, NJ), on the first day of STZ injections.

Techniques: Injection, Isolation, Incubation, Labeling