Overview
Type: Synthetic peptide
Form: Lyophilized powder
Orexin Receptor 1 Blocking Peptide (#BLP-OR001) is the original antigen used for immunization during Anti-Orexin Receptor 1 Antibody (#AOR-001) generation. The blocking peptide binds and ‘blocks’ Anti-Orexin Receptor 1 primary antibody, this makes it a good negative reagent control to help confirm antibody specificity in western blot and immunohistochemistry applications. This control is also often called a pre-adsorption control.
Applications: wb, ihc
For research purposes only. not for human use
Applications
Demonstration of Pre-adsorption control
Western blot analysis of mouse brain lysate:1. Anti-Orexin Receptor 1 Antibody (#AOR-001), (1:200).
2. Anti-Orexin Receptor 1 Antibody, preincubated with Orexin Receptor 1 Blocking Peptide (#BLP-OR001).
Western blot analysis of rat brain lysate:1. Anti-Orexin Receptor 1 Antibody (#AOR-001), (1:500).
2. Anti-Orexin Receptor 1 Antibody, preincubated with Orexin Receptor 1 Blocking Peptide (#BLP-OR001).
Western blot analysis of human Colo-205 (lanes 1 and 3) and HT-29 (lanes 2 and 4) colon cancer cell lines:1,2. Anti-Orexin Receptor 1 Antibody (#AOR-001), (1:500).
3,4. Anti-Orexin Receptor 1 Antibody, preincubated with Orexin Receptor 1 Blocking Peptide (#BLP-OR001).
Expression of OX1R in rat brainLongitudinal frozen section of rat brainstem showing staining (red) in neuronal cell bodies. Slides were incubated overnight at 4°C with Anti-Orexin Receptor 1 Antibody (#AOR-001), (1:50) followed by goat-anti-rabbit-AlexaFluor-555 secondary antibody (1:500). Hoechst 33342 is used as a counterstain (blue).
Expression of OX1R in rat colonImmunohistochemical staining of paraffin-embedded longitudinal section of rat colon showing mucosa (M), submucosa (SM), and muscularis externa (ME) using Anti-Orexin Receptor 1 Antibody (#AOR-001), (1:100). Note that the stain (red-brown color) is highly specific for absorptive cells in the superior third of the intestinal glands. Immunolabeling was detected using DAB as the chromogen and hematoxilin as the counterstain.
Expression of OX1R in mouse septumImmunohistochemical staining paraffin-fixed frozen sections using Anti-Orexin Receptor 1 Antibody (#AOR-001), (1:50). A. OX1R (green) appears in axonal processes (right-pointing triangles). B. Parvalbumin (red) appears in septal neurons. Cell nuclei (blue) are visualized with Hoechst 33342. C. Merge of OX1R and parvalbumin suggests that orexinergic innervation covers the entire septal nucleus rather than restricted to individual neurons.
Western blot analysis of rat brain membranes (lanes 1 and 3) and mouse brain lysate (lanes 2 and 4):1-2. Guinea Pig Anti-Orexin Receptor 1 Antibody (#AOR-001-GP), (1:200).
3-4. Guinea Pig Anti-Orexin Receptor 1 Antibody, preincubated with Orexin Receptor 1 Blocking Peptide (#BLP-OR001).
Expression of Orexin Receptor 1 in mouse locus coeruleus (LC)Immunohistochemical staining of perfusion-fixed frozen mouse brain sections with Guinea Pig Anti-Orexin Receptor 1 Antibody (#AOR-001-GP), (1:300), followed by goat anti-guinea pig-Alexa Fluor-594. A. OX1R immunoreactivity (red) appears in cells (vertical arrows) and in adjacent motor neurons of mesencephalic trigeminal nucleus (M5 horizontal arrows). B. Pre-incubation of the antibody with Orexin Receptor 1 Blocking Peptide (#BLP-OR001), suppressed staining. Cell nuclei are stained with DAPI (blue).
Expression of Orexin Receptor 1 in rat dorsal root nucleus (DRN).Immunohistochemical staining of perfusion-fixed frozen rat brain sections with Guinea Pig Anti-Orexin Receptor 1 Antibody (#AOR-001-GP), (1:300), followed by goat anti-guinea pig-Alexa Fluor-594. A. OX1R immunoreactivity (red) appears in DRN cells (arrows). B. Pre-incubation of the antibody with Orexin Receptor 1 Blocking Peptide (#BLP-OR001), suppressed staining. Cell nuclei are stained with DAPI (blue).
Properties
Sequence
- (C)RNWKRPSEQLEAQH, corresponding to amino acid residues 256-269 of rat OX1R (Accession P56718).
Accession (Uniprot) Number P56718
Peptide Confirmation Confirmed by amino acid analysis and mass spectrometry.
Purity >70%
Storage Before Reconstitution Lyophilized powder can be stored intact at room temperature for two weeks. For longer periods, it should be stored at -20°C.
Reconstitution 100 µl double distilled water (DDW).
Concentration After Reconstitution 0.4 mg/ml.
Storage After Reconstitution -20°C.
Antigen Preadsorption Control 1 μg peptide per 1 μg antibody.
Standard Quality Control Of Each Lot Western blot analysis.
