Recombinant Anti-LRP6 antibody [EPR22910-39] (ab231779)
![Western blot - Anti-LRP6 antibody [EPR22910-39] (ab231779)](https://www.abcam.com/ps/products/231/ab231779/Images/ab231779-512654-anti-lrp6-antibody-epr22910-39-western-blot-wildtype-hepg2-lrp6-knockout-hepg2-hela.jpg "Western blot - Anti-LRP6 antibody [EPR22910-39] (ab231779)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR22910-39] to LRP6
- Suitable for: WB
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-LRP6 antibody [EPR22910-39]
See all LRP6 primary antibodies -
Description
Rabbit monoclonal [EPR22910-39] to LRP6 -
Host species
Rabbit -
Tested applications
Suitable for: WBmore details
Unsuitable for: Flow Cyt,ICC/IF,IHC-P or IP -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HAP1 (WT), HeLa, HepG2 and HEK-293T whole cell lysates.
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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
EPR22910-39 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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KO cell lines
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab231779 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 |
1/1000. Detects a band of approximately 180-210 kDa (predicted molecular weight: 180 kDa).
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| Notes |
|---|
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WB
1/1000. Detects a band of approximately 180-210 kDa (predicted molecular weight: 180 kDa). |
Target
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Function
Component of the Wnt-Fzd-LRP5-LRP6 complex that triggers beta-catenin signaling through inducing aggregation of receptor-ligand complexes into ribosome-sized signalsomes. Cell-surface coreceptor of Wnt/beta-catenin signaling, which plays a pivotal role in bone formation. The Wnt-induced Fzd/LRP6 coreceptor complex recruits DVL1 polymers to the plasma membrane which, in turn, recruits the AXIN1/GSK3B-complex to the cell surface promoting the formation of signalsomes and inhibiting AXIN1/GSK3-mediated phosphorylation and destruction of beta-catenin. Required for posterior patterning of the epiblast during gastrulation. -
Tissue specificity
Widely co-expressed with LRP5 during embryogenesis and in adult tissues. -
Involvement in disease
Defects in LRP6 are the cause of autosomal dominant coronary artery disease type 2 (ADCAD2) [MIM:610947]. -
Sequence similarities
Belongs to the LDLR family.
Contains 4 EGF-like domains.
Contains 3 LDL-receptor class A domains.
Contains 20 LDL-receptor class B repeats. -
Domain
The YWTD-EGF-like domains 1 and 2 are required for the interaction with Wnt-frizzled complex. The YWTD-EGF-like domains 3 and 4 are required for the interaction with DKK1.
The PPPSP motifs play a central role in signal transduction by being phosphorylated, leading to activate the Wnt signaling pathway. -
Post-translational
modificationsDual phosphorylation of cytoplasmic PPPSP motifs sequentially by GSK3 and CK1 is required for AXIN1-binding, and subsequent stabilization and activation of beta-catenin via preventing GSK3-mediated phosphorylation of beta-catenin. Phosphorylated, in vitro, by GRK5/6 within and outside the PPPSP motifs. Phosphorylation at Ser-1490 by CDK14 during G2/M phase leads to regulation of the Wnt signaling pathway during the cell cycle. Phosphorylation by GSK3B is induced by RPSO1 binding and inhibited by DKK1. Phosphorylated, in vitro, by casein kinase I on Thr-1479.
Undergoes gamma-secretase-dependent regulated intramembrane proteolysis (RIP). The extracellular domain is first released by shedding, and then, through the action of gamma-secretase, the intracellular domain (ICD) is released into the cytoplasm where it is free to bind to GSK3B and to activate canonical Wnt signaling.
Palmitoylation on the two sites near the transmembrane domain leads to release of LRP6 from the endoplasmic reticulum.
Mono-ubiquitinated which retains LRP6 in the endoplasmic reticulum.
N-glycosylation is required for cell surface location. -
Cellular localization
Membrane. Endoplasmic reticulum. On Wnt signaling, undergoes a cycle of caveolin- or clathrin-mediated endocytosis and plasma membrane location. Released from the endoplasmic reticulum on palmitoylation. Mono-ubiquitination retains it in the endoplasmic reticulum in the absence of palmitoylation. On Wnt signaling, phosphorylated, aggregates and colocalizes with AXIN1 and GSK3B at the plasma membrane in LRP6-signalsomes. Chaperoned to the plasma membrane by MESD. - Information by UniProt
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Database links
- Entrez Gene: 4040 Human
- Omim: 603507 Human
- SwissProt: O75581 Human
- Unigene: 584775 Human
- Unigene: 658913 Human
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Alternative names
- ADCAD2 antibody
- C030016K15Rik antibody
- Cd antibody
see all
Images
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Western blot - Anti-LRP6 antibody [EPR22910-39] (ab231779)All lanes : Anti-LRP6 antibody [EPR22910-39] (ab231779) at 1/1000 dilution
Lane 1 : Wild-type HepG2 cell lysate
Lane 2 : LRP6 knockout HepG2 cell lysate
Lane 3 : HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 180 kDa
Observed band size: 180,200 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-LRP6 antibody [EPR22910-39] staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab231779 was shown to bind specifically to LRP6. A band was observed at 180/200 kDa in wild-type HepG2 cell lysates with no signal observed at this size in LRP6 knockout cell line ab277909 (knockout cell lysate ab284222). To generate this image, wild-type and LRP6 knockout HepG2 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
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![Western blot - Anti-LRP6 antibody [EPR22910-39] (ab231779)](https://www.abcam.com/ps/products/231/ab231779/Images/ab231779-352731-WB1anti-lrp6-antibody-epr-22910-39-western-blot-human.jpg)
Western blot - Anti-LRP6 antibody [EPR22910-39] (ab231779)All lanes : Anti-LRP6 antibody [EPR22910-39] (ab231779) at 1/1000 dilution
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : LRP6 knockout HAP1 whole cell lysate
Lane 3 : HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate
Lane 4 : HepG2 (human hepatocellular carcinoma epithelial cell), whole cell lysate
Lane 5 : 293T (human embryonic kidney epithelial cell), whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 180 kDa
Observed band size: 180-210 kDa why is the actual band size different from the predicted?The molecular weight observed is consistent with what has been described in the literature (PMID: 16989816, 16263759, 28341812). ab231779 was shown to specifically react with LRP6 in wild-type HAP1 cells as signal was lost in LRP6 knockout cells. Wild-type and LRP6 knockout samples were subjected to SDS-PAGE. ab231779 and ab129002 (Rabbit anti-Vinculin loading control) were incubated 1 hour at room temperature at 1/1000 dilution and 1/200,000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) secondary antibody at 1/20,000 dilution for 1 hour at room temperature before imaging. The blot was developed on a BIO-RAD® ChemiDoc™ instrument using the ECL technique. This blot was developed using a higher sensitivity ECL substrate.
Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure times: Lanes 1-2: 3 mins; Lanes 3: 26 secs; Lanes 4-5: 3 mins.
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![Western blot - Anti-LRP6 antibody [EPR22910-39] (ab231779)](https://www.abcam.com/ps/products/231/ab231779/Images/ab231779-355522-anti-lrp6-antibody-epr-22910-39-western-blot-human.jpg)
Western blot - Anti-LRP6 antibody [EPR22910-39] (ab231779)All lanes : Anti-LRP6 antibody [EPR22910-39] (ab231779) at 1/1000 dilution
Lane 1 : His-tagged human LRP5 recombinant protein, 10 ng
Lane 2 : His-tagged human LRP6 recombinant protein, 10 ng
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Predicted band size: 180 kDa
Observed band size: 18,36 kDa why is the actual band size different from the predicted?
Exposure time: 1 secondThe 36 KD band observed is consistent with the dimer of human LRP6 recombinant protein (aa 1476-1600).
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![Anti-LRP6 antibody [EPR22910-39] (ab231779)](https://www.abcam.com/ps/products/231/ab231779/Images/ab231779-3-benefits-of-recombinant-antibodies.png)
Anti-LRP6 antibody [EPR22910-39] (ab231779)
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 (0)
ab231779 has not yet been referenced specifically in any publications.