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4 protocols using goat anti mouse igg hrp

1

Western Blot Analysis of ER Stress Markers

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Sciatic nerves and RSC96 cells were lysed on ice in the RIPA buffer with protease inhibitor cocktail and phosphatase inhibitor cocktail for 30 min to extract total proteins. The proteins were analyzed with a bicinchoninic acid (BCA) protein assay kit (Biosynthesis, China). 30 μg/lane (sciatic nerve) and 20 μg/lane (RSC96 cell) were used for Western blot analysis as previously described8 (link),27 (link). The primary antibodies were as follows: mouse anti-IRE1α (sc-390960; 1: 1000; Santa Cruz), rabbit anti-P-IRE1α (ab48187; 1: 2000; abcam), mouse anti-XBP1 (sc-8015; 1: 1000; Abcam), mouse anti-GRP78 (sc-376768; 1: 1000; Santa Cruz), mouse anti-GADD153 (sc-7351; 1: 500; Santa Cruz), rabbit anti-Caspase-3 (ab13847; 1: 1000; abcam), rabbit anti-Caspase-12 (om273459; 1: 1000; Omnimabs), mouse anti-Bcl-2 (sc-7382; 1: 1000; Santa Cruz), mouse anti-Bax (sc-7480; 1: 1000; Santa Cruz), mouse anti-p-JNK (sc-6254; 1: 1000; Santa Cruz), and rabbit anti-PGP9.5 (ab108986; 1: 2000; Abcam). Mouse anti-β-actin (TA-09; 1: 20000; Zhongshan Goldenbridge) served as the internal control. The secondary antibodies were goat anti-mouse IgG-HRP (ZB-2305; Zhongshan Goldenbridge) and goat anti-rabbit IgG-HRP (ZB-2301; Zhongshan Goldenbridge). Western Chemiluminescent HRP Substrate and exposed to X-film to form image. The protein bands were quantitated with Image J software27 (link).
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2

Neuroprotective Mechanism of PEG-PCL Micelles

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Rosuvastatin was obtained from Solarbio (IR0150, Solarbio, Beijing, China). Poly (ethylene glycol) 2000-block-poly(ε-caprolactone) 3000 (PEG-PCL) was purchased from Shanghai ToYongBio Tech. Inc. (Shanghai, China). Fluoro-Jade C dyestuff was obtained from Merck (AG325-30MG, Millipore, Darmstadt, Germany). Rabbit anti-iNOS, rabbit anti-β-Tublin and mouse anti-Arginase 1 were purchased from Proteintech (1895-1-AP, 10094-1-AP, 66129-1-Ig, Proteintech, Wuhan, China). Goat anti-Iba-1 were purchased from Wako Pure Chemical Industries (011-27991, Wako, Osaka, Japan). Rabbit anti-IL-1β was supplied from Abbkine (ABP52932, Abbkine, Redlands, California). Rabbit anti- TNF-α was purchased from HuaAn Biotechnology (ER1919-22, HuaAnBio, Hangzhou, China). Rabbit anti- IL-10 was supplied from Wanleibio (WL03088, Wanleibio, Shenyang, China) Fluorescent Alexa 488 and 555 were purchased from Invitrogen (A-21121, A-11008, A-21432, Invitrogen, CA, USA). Peroxidase AffiniPure Goat Anti-Mouse IgG (H+L), AffiniPure Goat Anti-Rabbit IgG (H+L) were purchased from Jackson (115-035–003, 111-005-003, Jackson ImmunoResearch, PA, USA). The secondary antibodies for WB were goat anti-mouse IgG-HRP and goat anti-rabbit IgG-HRP (ZB-2301; ZB-2305; Zhongshan Golden Bridge, Beijing, China).
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3

Lycopene Modulates Angiogenic Factors

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Lycopene (purity ≥ 90%, from tomato), human fibronectin (FN), FITC-labeled Ulex, 1,1-dioctadecyl-3,3,3,3-tetramethylindocarbocyanine (Dil), labeled acetylated low-density lipoprotein (acLDL), and SB203580 were purchased from Sigma-Aldrich (St. Louis, MO, USA). M199 medium and fetal bovine serum were obtained from Hyclone (USA). Lydroxypropylmethyl Cellulose was purchased from Tianjin Haoyang Biological Manufacture Co., Ltd. China (Haoyang, China). RrhVEGF, recombinant human basic fibroblast growth factor (rhbFGF), epidermal growth factor (EGF), and insulin-like growth factor 1 (IGF-1) were purchased from Pepro Tech (USA). Annexin V-FITC Apoptosis Detection Kit was purchased from Jingmei Biotech Co., Ltd. (Shanghai, China). Boyden chamber was purchased from Haimen City Qilin Medical Instrument Factory. Antibodies against p38 MAPK (product code 9212) and phosphorylated p38 MAPK (product code 9216) were purchased from Cell Signaling Technology (Danvers, MA, USA). Goat Anti-mouse IgG/HRP (product code ZDR-5307) was purchased from Zhongshan Golden Bridge Bio-technology Co., Ltd. (Beijing, China). All other chemicals were obtained from common commercial sources and were of reagent grade. Pure Lycopene was dissolved in tetrahydrofuran (THF), stored at-70℃, protected from light [27 (link)], and diluted with medium to the desired concentration before use.
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4

Cytotoxicity Assay for Doxorubicin

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Doxorubicin hydrochloride, AG490, and bromodeoxyuridine (Brdu) were obtained from Sigma Aldrich (St. Louis, MO, USA). Dulbecco modified Eagle medium (DMEM) was obtained from Gibco (USA). Fetal bovine serum (FBS) was provided by Hyclone (Australia). The Hoechst 33258 staining kit was purchased from Beyotime Institute of Biotechnology (Shanghai, China). Lactate dehydrogenase (LDH) assay kit was provided by Biosino Bio-Technology and Science Inc (Beijing, China). Rabbit monoclonal anti-p-Stat3 (tyr-705) antibody, rabbit monoclonal anti-Stat3 (AB-727) antibody, rabbit monoclonal anti-p-Jak2 (tyr-1007) antibody, rabbit monoclonal anti-Jak2 (AB-1007) antibody, mouse anti-MT, and goat anti-rabbit immunoglobulin (Ig) G-horseradish peroxidase (HRP) were obtained from Sigma Aldrich (St. Louis, MO, USA). Mouse monoclonal anti-b-actin and goat anti-mouse IgG-HRP were purchased from Zhongshan Goldenbridge Biotechnology (Jiangsu, China).
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