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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-LRP5 antibody [EPR22477-218] - BSA and Azide free (ab256528)

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Western blot - Anti-LRP5 antibody [EPR22477-218] - BSA and Azide free (ab256528)
  • Immunoprecipitation - Anti-LRP5 antibody [EPR22477-218] - BSA and Azide free (ab256528)
  • Anti-LRP5 antibody [EPR22477-218] - BSA and Azide free (ab256528)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR22477-218] to LRP5 - BSA and Azide free
  • Suitable for: WB, IP
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

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HRP Conjugation Kit - Lightning-Link® (ab102890)
Protein
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Recombinant Human LRP5 protein (ab114399)

View more associated products

Overview

  • Product name

    Anti-LRP5 antibody [EPR22477-218] - BSA and Azide free
    See all LRP5 primary antibodies
  • Description

    Rabbit monoclonal [EPR22477-218] to LRP5 - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IPmore details
    Unsuitable for: Flow Cyt,ICC/IF or IHC-P
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Mouse LRP5 aa 1400 to the C-terminus. The exact sequence is proprietary.
    Database link: Q91VN0

  • Positive control

    • WB: SW260 and HEK293T cell lysates IP: 3T3-L1 whole cell lysate.
  • General notes

    ab256528 is the carrier-free version of ab223203 This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    Ab256528 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

     

    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
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    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

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C. Do Not Freeze.
  • Storage buffer

    pH: 7.2
    Constituent: PBS
  • Carrier free

    Yes
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR22477-218
  • Isotype

    IgG
  • Research areas

    • Cardiovascular
    • Lipids / Lipoproteins
    • Lipid Metabolism
    • Cholesterol Metabolism
    • Signal Transduction
    • Cytoskeleton / ECM
    • Extracellular Matrix
    • Structures
    • Bone
    • Neuroscience
    • Sensory System
    • Visual system
    • Stem Cells
    • Signaling Pathways
    • Wnt
    • Surface Molecules
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Lipid and lipoprotein metabolism
    • Cholesterol Metabolism

Associated products

  • Alternative Versions

    • Anti-LRP5 antibody [EPR22477-218] (ab223203)
  • Compatible Secondaries

    • VeriBlot for IP Detection Reagent (HRP) (ab131366)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human LRP5 knockout HEK293T cell line (ab266618)
  • KO cell lysates

    • Human LRP5 knockout HEK293T cell lysate (ab257202)
  • Recombinant Protein

    • Recombinant Human LRP5 protein (ab114399)

Applications

Our Abpromise guarantee covers the use of ab256528 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 at an assay dependent concentration. Detects a band of approximately 180-200 kDa (predicted molecular weight: 179 kDa).
IP Use at an assay dependent concentration.
  • Application notes
    Is unsuitable for Flow Cyt,ICC/IF or IHC-P.
  • Target

    • 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. Appears be required for postnatal control of vascular regression in the eye. Required for posterior patterning of the epiblast during gastrulation.
    • Tissue specificity

      Widely expressed, with the highest level of expression in the liver.
    • Involvement in disease

      Defects in LRP5 are the cause of vitreoretinopathy exudative type 4 (EVR4) [MIM:601813]. EVR4 is a disorder of the retinal vasculature characterized by an abrupt cessation of growth of peripheral capillaries, leading to an avascular peripheral retina. This may lead to compensatory retinal neovascularization, which is thought to be induced by hypoxia from the initial avascular insult. New vessels are prone to leakage and rupture causing exudates and bleeding, followed by scarring, retinal detachment and blindness. Clinical features can be highly variable, even within the same family. Patients with mild forms of the disease are asymptomatic, and their only disease related abnormality is an arc of avascular retina in the extreme temporal periphery. EVR4 inheritance can be autosomal dominant or recessive.
      Genetic variations in LRP5 are a cause of susceptibility to osteoporosis (OSTEOP) [MIM:166710]; also known as senile osteoporosis or postmenopausal osteoporosis. Osteoporosis is characterized by reduced bone mass, disruption of bone microarchitecture without alteration in the composition of bone. Osteoporotic bones are more at risk of fracture.
      Defects in LRP5 are the cause of osteoporosis-pseudoglioma syndrome (OPPG) [MIM:259770]; also known as osteogenesis imperfecta ocular form. OPPG is a recessive disorder characterized by very low bone mass and blindness. Individualy with OPPG are prone to develop bone fractures and deformations and have various eye abnormalities, including phthisis bulbi, retinal detachments, falciform folds or persistent vitreal vasculature.
      Defects in LRP5 are a cause of high bone mass trait (HBM) [MIM:601884]. HBM is a rare phenotype characterized by exceptionally dense bones. HBM individuals show otherwise a completely normal skeletal structure and no other unusual clinical findings.
      Defects in LRP5 are a cause of endosteal hyperostosis Worth type (WENHY) [MIM:144750]; also known as autosomal dominant osteosclerosis. WENHY is an autosomal dominant sclerosing bone dysplasia clinically characterized by elongation of the mandible, increased gonial angle, flattened forehead, and the presence of a slowly enlarging osseous prominence of the hard palate (torus palatinus). Serum calcium, phosphorus and alkaline phosphatase levels are normal. Radiologically, it is characterized by early thickening of the endosteum of long bones, the skull and of the mandible. With advancing age, the trabeculae of the metaphysis become thickened. WENHY becomes clinically and radiologically evident by adolescence, does not cause deformity except in the skull and mandible, and is not associated with bone pain or fracture. Affected patients have normal height, proportion, intelligence and longevity.
      Defects in LRP5 are the cause of osteopetrosis autosomal dominant type 1 (OPTA1) [MIM:607634]. Osteopetrosis is a rare genetic disease characterized by abnormally dense bone, due to defective resorption of immature bone. The disorder occurs in two forms: a severe autosomal recessive form occurring in utero, infancy, or childhood, and a benign autosomal dominant form occurring in adolescence or adulthood. OPTA1 is characterized by generalized osteosclerosis most pronounced in the cranial vault. Patients are often asymptomatic, but some suffer from pain and hearing loss. It appears to be the only type of osteopetrosis not associated with an increased fracture rate.
      Defects in LRP5 are the cause of van Buchem disease type 2 (VBCH2)[MIM:607636]. VBCH2 is an autosomal dominant sclerosing bone dysplasia characterized by cranial osteosclerosis, thickened calvaria and cortices of long bones, enlarged mandible and normal serum alkaline phosphatase levels.
    • 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.
    • Post-translational
      modifications

      Phosphorylation of cytoplasmic PPPSP motifs regulates the signal transduction of the Wnt signaling pathway through acting as a docking site for AXIN1.
    • Cellular localization

      Membrane. Endoplasmic reticulum. Chaperoned to the plasma membrane by MESD.
    • Target information above from: UniProt accession O75197 The UniProt Consortium
      The Universal Protein Resource (UniProt) in 2010
      Nucleic Acids Res. 38:D142-D148 (2010) .

      Information by UniProt
    • Database links

      • Entrez Gene: 4041 Human
      • Entrez Gene: 16973 Mouse
      • Entrez Gene: 293649 Rat
      • Omim: 603506 Human
      • SwissProt: O75197 Human
      • SwissProt: Q91VN0 Mouse
      • Unigene: 6347 Human
      • Unigene: 274581 Mouse
      • Alternative names

        • BMND1 antibody
        • EVR1 antibody
        • EVR4 antibody
        • HBM antibody
        • Low density lipoprotein receptor related protein 5 antibody
        • Low density lipoprotein receptor related protein 7 antibody
        • Low-density lipoprotein receptor-related protein 5 antibody
        • LR3 antibody
        • LRP-5 antibody
        • Lrp5 antibody
        • LRP5_HUMAN antibody
        • LRP7 antibody
        • OPPG antibody
        • OPS antibody
        • OPTA1 antibody
        • Osteoporosis pseudoglioma syndrome antibody
        • VBCH2 antibody
        see all

      Images

      • Western blot - Anti-LRP5 antibody [EPR22477-218] - BSA and Azide free (ab256528)
        Western blot - Anti-LRP5 antibody [EPR22477-218] - BSA and Azide free (ab256528)
        All lanes : Anti-LRP5 antibody [EPR22477-218] (ab223203) at 1/500 dilution

        Lane 1 : Wild-type HEK293T cell lysate
        Lane 2 : LRP5 knockout HEK293T cell lysate
        Lane 3 : SW620 cell lysate

        Lysates/proteins at 20 µg per lane.

        Secondary
        All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

        Predicted band size: 179 kDa
        Observed band size: 180-200 kDa
        why is the actual band size different from the predicted?



        This data was developed using the same antibody clone in a different buffer formulation (ab223203).

        Lanes 1-3: Merged signal (red and green). Green - ab223203 observed at 180-200 kDa. Red - loading control ab7291 observed at 50 kDa.

        ab223203 Anti-LRP5 antibody [EPR22477-218] was shown to specifically react with LRP5 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266618 (knockout cell lysate ab257202) was used. Wild-type and LRP5 knockout samples were subjected to SDS-PAGE. ab223203 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

         

      • Immunoprecipitation - Anti-LRP5 antibody [EPR22477-218] - BSA and Azide free (ab256528)
        Immunoprecipitation - Anti-LRP5 antibody [EPR22477-218] - BSA and Azide free (ab256528)

        LRP5 was immunoprecipitated from 0.35 mg 3T3-L1 (mouse embryonic fibroblast) differentiated adipocytes whole cell lysate 10µg with ab223203 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab223203. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/1000 dilution.

        Lane 1: 3T3-L1 (mouse embryonic fibroblast) differentiated adipocytes whole cell lysate 10µg.

        Lane 2: ab223203 IP in 3T3-L1 differentiated adipocytes whole cell lysate.

        Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab223203 in 3T3-L1 differentiated adipocytes whole cell lysate.

        Blocking/Dilution buffer: 5% NFDM/TBST.

        Exposure time: 3 mins.

        Differentiation procedure: https://www.abcam.com/protocols/differentiation-of-3t3-l1-cells-into-adipocyte-like-cells-protocol.

        This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab223203).

      • Anti-LRP5 antibody [EPR22477-218] - BSA and Azide free (ab256528)
        Anti-LRP5 antibody [EPR22477-218] - BSA and Azide free (ab256528)

      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.

      Click here to view the general protocols

      Datasheets and documents

      • Datasheet

      Certificate of Compliance

      To download a Certificate of Compliance, please enter your Lot number below:

    • References (0)

      Publishing research using ab256528? Please let us know so that we can cite the reference in this datasheet.

      ab256528 has not yet been referenced specifically in any publications.

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