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

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)
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STJ94837 Anti-Orexin R-1 antibody
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