Recombinant Anti-Orexin A antibody [EPR22803-259] (ab255294)
![Immunohistochemistry (Frozen sections) - Anti-Orexin A antibody [EPR22803-259] (ab255294)](https://www.abcam.com/ps/products/255/ab255294/Images/ab255294-5-anti-orexin-a-epr22803-259-immunohistochemistry-frozen-hypothalamus-rat.jpg "Immunohistochemistry (Frozen sections) - Anti-Orexin A antibody [EPR22803-259] (ab255294)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR22803-259] to Orexin A
- Suitable for: Dot blot, IHC-P, IHC-Fr, ELISA
- Reacts with: Mouse, Rat
Related conjugates and formulations
Overview
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Product name
Anti-Orexin A antibody [EPR22803-259]
See all Orexin A primary antibodies -
Description
Rabbit monoclonal [EPR22803-259] to Orexin A -
Host species
Rabbit -
Tested applications
Suitable for: Dot blot, IHC-P, IHC-Fr, ELISAmore details
Unsuitable for: Flow Cyt,ICC/IF or IP -
Species reactivity
Reacts with: Mouse, Rat -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- Western blot: His-tagged mouse Orexin A recombinant protein (aa33-130). Dot blot: Orexin A peptide (aa33-65). IHC-P: Mouse and rat hypothalamus tissue. IHC-Fr: Mouse and rat hypothalamus tissue.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR22803-259 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Immunohistochemistry kits
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab255294 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| Dot blot |
1/1000.
|
|
| IHC-P |
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
|
|
| IHC-Fr |
1/500.
Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20). |
|
| ELISA |
Use a concentration of 1 µg/ml.
|
| Notes |
|---|
|
Dot blot
1/1000. |
|
IHC-P
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
|
IHC-Fr
1/500. Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20). |
|
ELISA
Use a concentration of 1 µg/ml. |
Target
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Relevance
Orexin A (hypocretin 1) a 33 amino acid peptide and orexin B (hypocretin 2), a 28 amino acid peptide, are both derived from a common 130 amino acid precursor, prepro orexin. Orexin A and Orexin B stimulate food consumption when administered intracerebroventricularly to rats. Orexin gene expression in the brain is highly restricted to distinct populations of neurons located in specific hypothalamic regions, including the lateral hypothalamic area (LHA), a region implicated in feeding behaviour. Orexin A and orexin B bind to and activate two closely related G protein coupled receptors (GPCRs), termed Orexin 1 (OX1) and Orexin 2 (OX2) receptors. Dysfunction of the orexin peptide system has been linked to narcolepsy. -
Cellular localization
Rough endoplasmic reticulum. Cytoplasmic vesicle. Cell junction, synapse. Note: Associated with perikaryal rough endoplasmic reticulum as well as cytoplasmic large granular vesicles at synapses. -
Database links
- Entrez Gene: 15171 Mouse
- Entrez Gene: 25723 Rat
- SwissProt: O55241 Mouse
- SwissProt: O55232 Rat
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Alternative names
- HCRT antibody
- HCRT1 antibody
- HCRT2 antibody
see all
Images
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Immunohistochemistry (Frozen sections) - Anti-Orexin A antibody [EPR22803-259] (ab255294)
Immunohistochemical analysis of frozen section of 4% PFA-fixed, 0.2 % Triton X-100 permeabilized rat hypothalamus tissue labeling Orexin A with ab255294 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution (green). Positive staining on hypothalamic neurons (PMID: 9822755) is observed. The nuclear counterstain is DAPI (blue).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.
Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).
-
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Orexin A antibody [EPR22803-259] (ab255294)](https://www.abcam.com/ps/products/255/ab255294/Images/ab255294-3-anti-orexin-a-antibody-epr22803-259-immunohistochemistry-hypothalamus-rat.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Orexin A antibody [EPR22803-259] (ab255294)Immunohistochemical analysis of paraffin-embedded rat hypothalamus tissue labeling Orexin A with ab255294 at 1/1000 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining in rat hypothalamus (PMID: 9419374) is observed. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
The section was incubated with ab255294 for 15 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
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![Dot Blot - Anti-Orexin A antibody [EPR22803-259] (ab255294)](https://www.abcam.com/ps/products/255/ab255294/Images/ab255294-1-anti-orexin-a-antibody-epr22803-259-dot-mouse.jpg)
Dot Blot - Anti-Orexin A antibody [EPR22803-259] (ab255294)Dot blot analysis of Orexin A labeled with ab255294 at 1/1000 dilution.
Lane 1: Mouse Orexin A peptide (aa33-65).
Lane 2: Mouse Orexin B peptide (aa69-96).Blocking and dilution buffer: 5% NFDM/TBST.
Exposure time: 3 minutes.
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![ELISA - Anti-Orexin A antibody [EPR22803-259] (ab255294)](https://www.abcam.com/ps/products/255/ab255294/Images/ab255294-352157-anti-orexin-a-antibody-epr22803259-elisa.jpg)
ELISA - Anti-Orexin A antibody [EPR22803-259] (ab255294)Direct ELISA antigen dose-response curve using ab255294 at 1 µg/mL. Antigen (mouse Orexin A peptide) concentration of 0.488-500 ng/mL. ab97048 Goat Anti-Rabbit IgG H&L (Alkaline Phosphatase) at 1/5000 was used as the secondary antibody.
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![Immunohistochemistry (Frozen sections) - Anti-Orexin A antibody [EPR22803-259] (ab255294)](https://www.abcam.com/ps/products/255/ab255294/Images/ab255294-4-anti-orexin-a-epr22803-259-immunohistochemistry-frozen-hypothalamus-mouse.jpg)
Immunohistochemistry (Frozen sections) - Anti-Orexin A antibody [EPR22803-259] (ab255294)Immunohistochemical analysis of frozen section of 4% PFA-fixed, 0.2 % Triton X-100 permeabilized mouse hypothalamus tissue labeling Orexin A with ab255294 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution (green). Positive staining on hypothalamic neurons (PMID: 9822755) is observed. The nuclear counterstain is DAPI (blue).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.
Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).
-
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Orexin A antibody [EPR22803-259] (ab255294)](https://www.abcam.com/ps/products/255/ab255294/Images/ab255294-2-anti-orexin-a-antibody-epr22803-259-immunohistochemistry-hypothalamus-mouse.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Orexin A antibody [EPR22803-259] (ab255294)Immunohistochemical analysis of paraffin-embedded mouse hypothalamus tissue labeling Orexin A with ab255294 at 1/1000 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining in mouse hypothalamus (PMID: 9419374) is observed. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
The section was incubated with ab255294 for 15 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
-
![Anti-Orexin A antibody [EPR22803-259] (ab255294)](https://www.abcam.com/ps/products/255/ab255294/Images/ab255294-8-benefits-of-recombinant-antibodies.png)
Anti-Orexin A antibody [EPR22803-259] (ab255294)
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (1)
ab255294 has been referenced in 1 publication.
- Gargiulo AT et al. Sex differences in cognitive flexibility are driven by the estrous cycle and stress-dependent. Front Behav Neurosci 16:958301 (2022). PubMed: 35990724