goat antihuman lepr  (Bio-Rad)

Journal: The Journal of clinical endocrinology and metabolism

Article Title: Estradiol affects extracellular leptin:adiponectin ratio in human breast tissue in vivo.

doi: 10.1210/jc.2014-1129

Figure Lengend Snippet: Figure 3. Tamoxifen significantly decreased leptin and increased adiponectin in normal human breast tissue. A. Postmenopausal women underwent microdialysis sampling of breast tissue (n 18) and subcutaneous (sc) abdominal fat (n 16) before tamoxifen treatment and 6 weeks after treatment. *P .05, **P .01. B. To confirm the in vivo findings, whole normal human breast tissue biopsies were cultured with or without 10-6 M tamoxifen (Tam) for 7 days, and adipokine levels in the culture medium were determined. *P .05 (n 9), ****P .0001 (n 12) compared to controls. C. Normal breast tissue biopsies were cultured ex vivo with or without 10-6 M tamoxifen (Tam). Levels of adipokines and their corresponding receptors (leptin receptor [LepR], adiponectin receptor 1 [AdipoR1], and adiponectin receptor 2 [AdipoR2]) were determined by immunohistochemistry analysis. No differences were detected between treatment groups. Representative tissue sections from each treatment group are shown (scale bars, 20 m).

Article Snippet: Immunohistochemistry Formalin-fixed, paraffin-embedded normal breast tissue biopsies and mouse tumors were cut into 4- m sections, deparaffinized, and subjected to high-temperature antigen retrieval prior to incubating with the following primary antibodies: rabbit antihuman leptin (1 g/ml; GenWay Biotech, San Diego, CA, USA), goat antihuman LepR (20 g/ml; AbD Serotec, Düsseldorf, Germany), goat antimouse leptin (10 g/ml; R&D Systems), goat antimouse LepR (10 g/ml; R&D Systems), rabbit antihuman adiponectin (1 g/ml; Abcam, Cambridge, UK), rabbit antihuman AdipoR1 (5 g/ml; Bioss, London, UK), and rabbit antihuman AdipoR2 (5 g/ml; Bioss).

Techniques: Sampling, In Vivo, Cell Culture, Ex Vivo, Immunohistochemistry

Journal: The Journal of clinical endocrinology and metabolism

Article Title: Estradiol affects extracellular leptin:adiponectin ratio in human breast tissue in vivo.

doi: 10.1210/jc.2014-1129

Figure Lengend Snippet: Figure 4. Increased extracellular leptin and decreased adiponectin in human breast cancer in vivo. A. Eleven postmenopausal breast cancer patients underwent microdialysis sampling of the tumor and normal adjacent breast tissue before surgery. *P .05, **P .01. B. FVB/N mice were oophorectomized and treated with or without estradiol (physiologic level), and MMTV- PyMT mammary cancer cells were injected into the dorsal mammary fat pad. At similar tumor sizes, both groups of mice underwent microdialysis sampling of adipokines in the tumor. *P .05, **P .01 (each group, n 7). C. FVB/N mice were treated as described in Figure 4B. Levels of adipokines and their corresponding receptors (leptin receptor [LepR], adiponectin receptor 1 [AdipoR1], and adiponectin receptor 2 [AdipoR2]) in the MMTV-PyMT tumors were determined by immunohistochemistry analysis. Increased LepR staining was observed in estrogen-exposed tumors. Representative tissue sections from each treatment group are shown (scale bars, 20 m).

Article Snippet: Immunohistochemistry Formalin-fixed, paraffin-embedded normal breast tissue biopsies and mouse tumors were cut into 4- m sections, deparaffinized, and subjected to high-temperature antigen retrieval prior to incubating with the following primary antibodies: rabbit antihuman leptin (1 g/ml; GenWay Biotech, San Diego, CA, USA), goat antihuman LepR (20 g/ml; AbD Serotec, Düsseldorf, Germany), goat antimouse leptin (10 g/ml; R&D Systems), goat antimouse LepR (10 g/ml; R&D Systems), rabbit antihuman adiponectin (1 g/ml; Abcam, Cambridge, UK), rabbit antihuman AdipoR1 (5 g/ml; Bioss, London, UK), and rabbit antihuman AdipoR2 (5 g/ml; Bioss).

Techniques: In Vivo, Sampling, Injection, Immunohistochemistry, Staining