Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-LRP5 antibody [EPR22477-218] (ab223203)

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
  • Suitable for: WB, IP
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-LRP5 antibody [EPR22477-218]
    See all LRP5 primary antibodies

  • Description

    Rabbit monoclonal [EPR22477-218] to LRP5

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    IP
    Mouse
    WB
    Mouse
    Rat
    Human
    See all applications and species data

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: MCF7, HCT 116, MEF, 3T3-L1 and HeLa whole cell lysates; rat liver tissue lysate; His-tagged mouse LRP5 recombinant protein (aa 1407-1614). IP: 3T3-L1 whole cell lysate.

  • 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
    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

Images

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

    Lane 1 : Wild-type HEK-293T cell lysate
    Lane 2 : LRP5 knockout HEK293T cell lysate
    Lane 3 : SW620 cell lysate

    Blocking peptides 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?



    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.

     

  • Western blot - Anti-LRP5 antibody [EPR22477-218] (ab223203)
    Western blot - Anti-LRP5 antibody [EPR22477-218] (ab223203)
    All lanes : Anti-LRP5 antibody [EPR22477-218] (ab223203) at 1/1000 dilution

    Lane 1 : MCF7 (human breast adenocarcinoma epithelial cell), whole cell lysate
    Lane 2 : HCT116 (human colorectal carcinoma epithelial cell), whole cell lysate
    Lane 3 : HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate
    Lane 4 : Rat liver tissue 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: 179 kDa


    Exposure time: 3 minutes


    The expression profile observed is consistent with what has been described in the literature (PMID: 25808845).

    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-LRP5 antibody [EPR22477-218] (ab223203)
    Western blot - Anti-LRP5 antibody [EPR22477-218] (ab223203)
    All lanes : Anti-LRP5 antibody [EPR22477-218] (ab223203) at 1/1000 dilution

    Lane 1 : MEF (mouse embryonic fibroblast (immortalized)), whole cell lysate at 20 µg
    Lane 2 : Untreated 3T3-L1 (mouse embryonic fibroblast), whole cell lysate at 20 µg
    Lane 3 : 3T3-L1 differentiated adipocytes whole cell lysate 20ug

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

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



    The expression profile observed is consistent with what has been described in the literature (PMID: 25808845).

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

    Exposure times: Lane 1: 3 mins; Lane 2-3: 26 secs.

    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-LRP5 antibody [EPR22477-218] (ab223203)
    Western blot - Anti-LRP5 antibody [EPR22477-218] (ab223203)
    All lanes : Anti-LRP5 antibody [EPR22477-218] (ab223203) at 1/1000 dilution

    Lane 1 : His-tagged mouse LRP5 recombinant protein (aa 1407-1614)
    Lane 2 : His-tagged mouse LRP6 recombinant protein (aa 1394-1613)

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 179 kDa
    Observed band size: 25.1,37.5 kDa why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: 5.5 secs.

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

    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.

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

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

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