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Western Blot Antibody Customer Review for Anti-Orexin R-1 Polyclonal Antibody (STJ94837)

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The Oxerin receptor type 1 protein (also known as hypocretin receptor type 1) is a G protein coupled receptor mainly expressed in the hypothalamus. It is thought to be involved in feeding behaviour. The protein is encoded by the HCRTR1 gene in humans. The Orexin R-1 polyclonal antibody detects endogenous levels of Orexin R-1 protein.

Anti-Orexin R-1 antibody - http://www.stjohnslabs.com/orexin-r-1-antibody-p-93702?filter_name=STJ94837

Join Our Antibody Validation Project - http://www.stjohnslabs.com/services/antibody-validation

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Western Blot Antibody Customer Review for Anti-Orexin R-1 Polyclonal Antibody (STJ94837)

  1. 1. Result “In mouse striatal progenitors target bands were detected by STJ94837 Orexin R-1 Antibody. Two bands were detected – ~65 and ~80 kDa. The molecular weight of orexin R-1 is 50 kDa.” - K. Palm, Protobios. Gel electrophoresis information:10% polyacrylamide gel, electric current 35 mA, 90 minutes. Transfer information:0.45 um PVDF membrane, 1 piece of gel (wet transfer), electric current 350 mA 95 minutes. Lane No. Antigen Loading amount Primary antibody Primary antibody dilution ratio Secondary antibody dilution ratio Target band KD Visualizati on time 1 Striatal progenitors 10 ug STJ94837 Anti- Orexin R-1 antibody 1:1000 1:10000 ~50KD 5 minutes Cell Line: Mouse striatal progenitors Method of validation: Western Blot Primary Antibody: STJ94837 Anti-Orexin R-1 antibody Secondary Antibody HRP goat anti-rabbit Reference antibody: GAPTH (Sigma) Dilution ratio: 1:1000 Protocol Treatment of materials: Cells were cultured to 70%-80% plating density, cytoplasmic fraction was extracted by hypotonic buffer (10 mM Hepes, 1,5 mM MgCl, 10 mM KCL, 0,5 mM DTT, ph 7,9) with protease inhibitors, 5x loading buffer was added, denaturation was performed at 50 ˚C for 10 minutes. Gel electrophoresis: 10% polyacrylamide gel, electric current 35 mA, 90 minutes. Transfer: 0.45 um PVDF membrane, 1 piece of gel (wet transfer), electric current 350 mA 95 minutes. Blocking: 1X TBS Tween (0,1%) with 5% skimmed milk powder for 1 hour. Primary antibody probing: primary antibody was added to the solution of 1X TBS Tween (0,1%) with 5% skimmed milk powder, 4˚C for overnight. Membrane wash: 1X TBS Tween (0.1%) wash for 3 times. Secondary antibody probing: secondary antibody was added to the solution of 1X TBS Tween (0,1%) with 5% skimmed milk powder with 1:10000 dilution ratio, for 1 hour. Membrane wash: 1X TBS Tween (0.1%) wash for 3 times. Visualization: ECL visualization for 5 minutes with SuperSignal West Pico ECL-kit (Thermo scientific) Antibody Customer Review: STJ94837 Anti-Orexin R-1 antibody Antibody Specificity: Antibody Rating:

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