Validated using a knockout cell line
Recombinant
RabMAb

Anti-CD168 antibody [EPR4054] (ab124729)

Overview

  • Product name
    Anti-CD168 antibody [EPR4054]
    See all CD168 primary antibodies
  • Description
    Rabbit monoclonal [EPR4054] to CD168
  • Tested applications
    Suitable for: WB, IP, IHC-Pmore details
    Unsuitable for: Flow Cyt or ICC
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    corresponding to Human CD168 aa 1-100.

  • Positive control
    • Lysates of: MDA-MB-435, LnCaP, T47-D, RAW264.7, PC-12. Human testis tissue
  • General notes

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

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab124729 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. Predicted molecular weight: 84 kDa.

For unpurified use at 1/1000 - 1/10000.

The mouse and rat recommendation is based on the WB results. This antibody may not be suitable for IHC with mouse or rat samples.

IP 1/10 - 1/100.
IHC-P 1/250.

For unpurified use at 1/50 - 1/100.

Antigen retrieval is recommended

  • Application notes
    Is unsuitable for Flow Cyt or ICC.
  • Target

    • Function
      Involved in cell motility. When hyaluronan binds to HMMR, the phosphorylation of a number of proteins, including the focal adhesion kinase occurs. May also be involved in cellular transformation and metastasis formation, and in regulating extracellular-regulated kinase (ERK) activity.
    • Tissue specificity
      Expressed in breast cancer cell lines and in normal breast tissue.
    • Cellular localization
      Cell surface. Cytoplasm.
    • Information by UniProt
    • Database links
    • Alternative names
      • CD168 antibody
      • CD168 antigen antibody
      • HMMR antibody
      • HMMR_HUMAN antibody
      • Hyaluronan mediated motility receptor antibody
      • Hyaluronan-mediated motility receptor (RHAMM) antibody
      • IHABP antibody
      • Intracellular hyaluronic acid-binding protein antibody
      • MGC119494 antibody
      • MGC119495 antibody
      • OTTHUMP00000196920 antibody
      • Receptor for hyaluronan-mediated motility antibody
      • RHAMM antibody
      see all

    Anti-CD168 antibody [EPR4054] images

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human gastric carcinoma tissue sections labeling CD168 with purified  ab124729 at 1:250 dilution (7.7 μg/ml). Heat mediated antigen retrieval was performed using Perform heat mediated antigen retrieval using EDTA Buffer, pH9.0. Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.

    • ab124729 (purified) at 1:100 dilution (2ug) immunoprecipitating CD168 in MCF-7 (Human breast adenocarcinoma epithelial cell) whole cell lysate.

      Lane 1 (input): MCF-7 (Human breast adenocarcinoma epithelial cell) whole cell lysate 10ug

      Lane 2 (+): ab124729 & MCF-7 (Human breast adenocarcinoma epithelial cell) whole cell lysate

      Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab124729 in MCF-7 (Human breast adenocarcinoma epithelial cell) whole cell lysate

      For western blotting, VeriBlot for IP secondary antibody (HRP) (ab131366) was used as the secondary antibody at 1:1000 dilution.

      Blocking and diluting buffer: 5% NFDM/TBST.



    • Predicted band size : 84 kDa

      Lane 1: Wild type HAP1 whole cell lysate (20 µg)
      Lane 2: Empty knockout  HAP1 whole cell lysate (20 µg)
      Lane 3: CD168 whole cell lysate (20 µg)

      Lanes 1 - 3: Merged signal (red and green). Green - ab124729 observed at 90 kDa. Red - loading control, ab9484, observed at 37 kDa.

      ab124729 was shown to specifically react with CD168 when CD168 knockout samples were used. Wild-type and Empty knockout samples were subjected to SDS-PAGE.  Ab124729 and ab9484 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1000 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

    • Immunohistochemical analysis of paraffin-embedded human testis tissue using  unpurified ab124729 at 1/50 - 1/100 dilution

    • All lanes : Anti-CD168 antibody [EPR4054] (ab124729) at 1/1000 dilution (purified)

      Lane 1 : MDA-MB-435 (Human mammary gland ductal carcinoma melanocyte) whole cell lysates at 20 µg
      Lane 2 : RAW264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysates at 15 µg
      Lane 3 : PC-12 (Rat adrenal gland pheochromocytoma ) whole cell lysates at 15 µg

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size : 84 kDa

      5% NFDM/TBST

    • All lanes : Anti-CD168 antibody [EPR4054] (ab124729)

      Lane 1 : MDA-MB-435 cell lysate
      Lane 2 : LnCaP cell lysate
      Lane 3 : T47-D cell lysate

      Lysates/proteins at 10 µg per lane.


      Predicted band size : 84 kDa
    • Equilibrium disassociation constant (KD)
      Learn more about KD

      Click here to learn more about KD

    References for Anti-CD168 antibody [EPR4054] (ab124729)

    This product has been referenced in:
    • Song JM  et al. Triptolide suppresses the in vitro and in vivo growth of lung cancer cells by targeting hyaluronan-CD44/RHAMM signaling. Oncotarget 8:26927-26940 (2017). WB ; Human . Read more (PubMed: 28460475) »
    • Coulson-Thomas VJ  et al. Umbilical Cord Mesenchymal Stem Cells Suppress Host Rejection: THE ROLE OF THE GLYCOCALYX. J Biol Chem 289:23465-81 (2014). ICC/IF ; Human . Read more (PubMed: 24986866) »

    See all 2 Publications for this product

    Product Wall

    Application
    Western blot
    Loading amount
    35 µg
    Gel Running Conditions
    Reduced Denaturing (10% SDS-PAGE)
    Sample
    Dog Cell lysate - whole cell (canine mammay tumor cells)
    Specification
    canine mammay tumor cells
    Treatment
    siRNA against CD168 for 24 hrs
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Username

    光芳 齋藤 

    Verified customer

    Submitted Jan 20 2015

    For ab124729 lot GR82264-8, the concentration is 1.797 mg/ml.

    The current lot we have for sale for ab108339 is lot GR106937-4 and that is 0.193 mg/ml.

    Thank you for contacting Abcam earlier today.

    Please find attached the IHC-P protocol for both ab108339 & ab124729.

    If there is anything else I can help you with, please let me know.

    Thank you for your email.



    I am glad to hear that the free of charge replacement worked as expected and you have obtained satisfactory results.



    If you wish, you could submit your results as feedback using our Ab...

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