Anti-Leptin Receptor antibody (ab104403)
Key features and details
- Rabbit polyclonal to Leptin Receptor
- Suitable for: WB, IHC-P, ICC/IF
- Reacts with: Human
- Isotype: IgG
Overview
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Product name
Anti-Leptin Receptor antibody
See all Leptin Receptor primary antibodies -
Description
Rabbit polyclonal to Leptin Receptor -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-P, ICC/IFmore details -
Species reactivity
Reacts with: Human
Predicted to work with: Macaque monkey
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Immunogen
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Positive control
- This antibody gave a positive signal in HepG2 whole cell lysate. This antibody gave a positive result in IHC in the following FFPE tissue: human normal kidney.
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General notes
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
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 or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration
Concentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
Our Abpromise guarantee covers the use of ab104403 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 WB Use a concentration of 1 µg/ml. Detects a band of approximately 150 kDa (predicted molecular weight: 132 kDa). IHC-P Use a concentration of 5 µg/ml. ICC/IF Use a concentration of 1 µg/ml. Target
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Function
Receptor for obesity factor (leptin). On ligand binding, mediates signaling through JAK2/STAT3. Involved in the regulation of fat metabolism and, in a hematopoietic pathway, required for normal lymphopoiesis. May play a role in reproduction. Can also mediate the ERK/FOS signaling pathway. -
Tissue specificity
Isoform A is expressed in fetal liver and in hematopoietic tissues and choroid plexus. In adults highest expression in heart, liver, small intestine, prostate and ovary. Low level in lung and kidney. Isoform B is highly expressed in hypothalamus. -
Sequence similarities
Belongs to the type I cytokine receptor family. Type 2 subfamily.
Contains 4 fibronectin type-III domains.
Contains 1 Ig-like (immunoglobulin-like) domain. -
Domain
The cytoplasmic domain may be essential for intracellular signal transduction by activation of JAK tyrosine kinase and STATs.
The WSXWS motif appears to be necessary for proper protein folding and thereby efficient intracellular transport and cell-surface receptor binding.
The box 1 motif is required for JAK interaction and/or activation. -
Post-translational
modificationsOn ligand binding, phosphorylated on two conserved C-terminal tyrosine residues (isoform B only) by JAK2. Tyr-986 is required for complete binding and activation of PTPN11, ERK/FOS activation and, for interaction with SOCS3 (By similarity). Phosphorylation on Tyr-1141 is required for STAT3 binding/activation. -
Cellular localization
Secreted and Cell membrane. - Information by UniProt
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Database links
- Entrez Gene: 3953 Human
- Omim: 601007 Human
- SwissProt: P48357 Human
- Unigene: 23581 Human
- Unigene: 723178 Human
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Alternative names
- CD 295 antibody
- CD295 antibody
- CD295 antigen antibody
see all
Images
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Western blot - Anti-Leptin Receptor antibody (ab104403)Anti-Leptin Receptor antibody (ab104403) at 1 µg/ml + HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate at 10 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 132 kDa
Observed band size: 150 kDa why is the actual band size different from the predicted?
Additional bands at: 40 kDa, 49 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 1 minute
Leptin Receptor contains an exstensive number of potential glycosylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted. -
Immunocytochemistry/ Immunofluorescence - Anti-Leptin Receptor antibody (ab104403)ICC/IF image of ab104403 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab104403 at 1µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Leptin Receptor antibody (ab104403)IHC image of aLeptin Receptor staining in Human normal kidney formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab104403, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
Protocols
References (11)
ab104403 has been referenced in 11 publications.
- Gao B et al. Leptin receptor-expressing cells represent a distinct subpopulation of notochord-derived cells and are essential for disc homoeostasis. J Orthop Translat 21:91-99 (2020). PubMed: 32110508
- Nogues P et al. Maternal obesity influences expression and DNA methylation of the adiponectin and leptin systems in human third-trimester placenta. Clin Epigenetics 11:20 (2019). PubMed: 30732639
- Kong Y et al. The Effect of the Leptin and Leptin Receptor Expression on the Efficacy of Neoadjuvant Chemotherapy in Breast Cancer. Med Sci Monit 25:3005-3013 (2019). PubMed: 31015393
- Al-Shibli SM et al. Expression of leptin and leptin receptors in colorectal cancer-an immunohistochemical study. PeerJ 7:e7624 (2019). PubMed: 31592340
- Chavira-Suárez E et al. Influence of pre-pregnancy body mass index (p-BMI) and gestational weight gain (GWG) on DNA methylation and protein expression of obesogenic genes in umbilical vein. PLoS One 14:e0226010 (2019). PubMed: 31794592
Images
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Western blot - Anti-Leptin Receptor antibody (ab104403)Anti-Leptin Receptor antibody (ab104403) at 1 µg/ml + HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate at 10 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 132 kDa
Observed band size: 150 kDa why is the actual band size different from the predicted?
Additional bands at: 40 kDa, 49 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 1 minute
Leptin Receptor contains an exstensive number of potential glycosylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted. -
Immunocytochemistry/ Immunofluorescence - Anti-Leptin Receptor antibody (ab104403)
ICC/IF image of ab104403 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab104403 at 1µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Leptin Receptor antibody (ab104403)
IHC image of aLeptin Receptor staining in Human normal kidney formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab104403, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
