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2 protocols using mouse antihuman monoclonal antibodies

1

Granulosa Cell Culture Protocols

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All cell culture materials were obtained from Gibco Inc. Recombinant Human Activin-A (GFH6) was purchased from Cell Guidance Systems. RepSox (#72392), a cell permeable, selective inhibitor of the TGF-β type 1 receptor (TGFβRI) ALK5 was obtained from Stemcell Technologies. Recombinant forms of FSH (Gonal-F) and hCG (Ovitrelle) was purchased from Merck Global (Darmstadt, Germany). SAPK/JNK Kinase Assay Kit (#8794, nonradioactive), Hoechst 33342 (#4082), anti c-Jun (#9165), anti-phospho-c-JunSer73 (#3270S), Anti-Phospho-SAPK/JNKThr183/Tyr185 (#9251), Smad2 (#3122) and Phospho-Smad2Ser465/Ser467 (#18338) antibodies were obtained from Cell Signaling. Oil Red O was purchased from Sigma Inc. (USA). All western blotting buffers and reagents were purchased from Bio-Rad. Anti-Vinculin Antibody (V9264) was purchased from Sigma-Aldrich. Mouse antihuman monoclonal antibodies were purchased from Santa Cruz Biotechnology for the detection of human 3β-HSD Type II (sc-100466), 17β-HSD type-I (sc-376719), StAR (sc-166821), and P450 SCC (CYP11A1, sc-292456). Aromatase (CYP19A1, ab34193) monoclonal mouse antibody was from Abcam Inc. YO-PRO®-1 Iodide (491/509) was obtained from Life Technologies. Texas Red™-X Phalloidin was obtained from Thermo Fisher Scientific.
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2

Immunoblotting Assay for MIC A/B and NKG2D

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TRIzol was purchased from Invitrogen Life Technologies (Carlsbad, CA, USA) and M-MLV reverse transcriptase was purchased from Promega Corporation (Madison, WI, USA). MIC A/B and NKG2D mouse anti-human monoclonal antibodies were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA), hypoxia-inducible factor 1-α (HIF-1α) rabbit anti-human polyclonal antibody was purchased from Boster Systems Inc. (Pleasanton, CA, USA) and glyceryl trinitrate (GTN) transdermal patches were purchased from Schwarz Pharma (Brussels, Belgium).
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