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  1. Link

    cd168-antibody-epr4055-ab108339.pdf

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Stem Cells Embryonic Stem Cells Surface Molecules
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-CD168 antibody [EPR4055] (ab108339)

  • Datasheet
  • SDS
Submit a review Q&A (6)References (4)

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Western blot - Anti-CD168 antibody [EPR4055] (ab108339)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD168 antibody [EPR4055] (ab108339)
  • Immunoprecipitation - Anti-CD168 antibody [EPR4055] (ab108339)
  • Western blot - Anti-CD168 antibody [EPR4055] (ab108339)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD168 antibody [EPR4055] (ab108339)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD168 antibody [EPR4055] (ab108339)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD168 antibody [EPR4055] (ab108339)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD168 antibody [EPR4055] (ab108339)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD168 antibody [EPR4055] (ab108339)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD168 antibody [EPR4055] (ab108339)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD168 antibody [EPR4055] (ab108339)
  • Anti-CD168 antibody [EPR4055] (ab108339)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR4055] to CD168
  • Suitable for: WB, IP, IHC-P
  • Knockout validated
  • Reacts with: Human

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Overview

  • Product name

    Anti-CD168 antibody [EPR4055]
    See all CD168 primary antibodies
  • Description

    Rabbit monoclonal [EPR4055] to CD168
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IP, IHC-Pmore details
    Unsuitable for: ICC/IF
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • T47-D MCF-7, SKBR-3 and LnCaP cell lysate Paraffin-embedded human breast carcinoma tissue Paraffin-embedded human testis tissue IP: T-47D 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.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
  • Storage buffer

    pH: 7.20
    Preservative: 0.05% Sodium azide
    Constituents: 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR4055
  • Isotype

    IgG
  • Research areas

    • Stem Cells
    • Embryonic Stem Cells
    • Surface Molecules
    • Cancer
    • Tumor immunology
    • CD markers
    • Developmental Biology
    • Embryogenesis
    • Embryonic stem cells
    • Intracellular

Associated products

  • Alternative Versions

    • Anti-CD168 antibody [EPR4055] - Low endotoxin, Azide free (ab229447)
    • Anti-CD168 antibody [EPR4055] - BSA and Azide free (ab271875)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
    • Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • T-47D whole cell lysate (ab14899)
    • MCF7 whole cell lysate (ab29537)
    • MCF7 membrane extract lysate (ab29539)
  • Recombinant Protein

    • Recombinant Human CD168 protein (ab125997)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab108339 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 - 1/10000. Predicted molecular weight: 84 kDa.
IP
1/10 - 1/100.
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Notes
WB
1/1000 - 1/10000. Predicted molecular weight: 84 kDa.
IP
1/10 - 1/100.
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Application notes
Is unsuitable for ICC/IF.

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.
  • Target information above from: UniProt accession O75330 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 3161 Human
    • Omim: 600936 Human
    • SwissProt: O75330 Human
    • Unigene: 728200 Human
    • 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

    Images

    • Western blot - Anti-CD168 antibody [EPR4055] (ab108339)
      Western blot - Anti-CD168 antibody [EPR4055] (ab108339)

      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 - ab108339 observed at 90 kDa. Red - loading control, ab9484, observed at 37 kDa.

      ab108339 was shown to specifically react with CD168 when CD168 knockout samples were used. Wild-type and Empty knockout samples were subjected to SDS-PAGE.  Ab108339 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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD168 antibody [EPR4055] (ab108339)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD168 antibody [EPR4055] (ab108339)

      ab108339, at 1/100, staining CD168 in paraffin-embedded human breast carcinoma tissue by Immunohistochemistry.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • Immunoprecipitation - Anti-CD168 antibody [EPR4055] (ab108339)
      Immunoprecipitation - Anti-CD168 antibody [EPR4055] (ab108339)

      Purified ab108339 at 1/50 dilution (2µg) immunoprecipitating CD168 in T-47D whole cell lysate.
      Lane 1 (input): T-47D (Human ductal breast epithelial tumor epithelial cell) whole cell lysate 10µg
      Lane 2 (+): ab108339 + T-47D whole cell lysate.
      Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab108339 in T-47D whole cell lysate.
      VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
      Blocking Buffer and concentration: 5% NFDM/TBST.
      Diluting buffer and concentration: 5% NFDM/TBST.
      Observed band size: 85 kDa

    • Western blot - Anti-CD168 antibody [EPR4055] (ab108339)
      Western blot - Anti-CD168 antibody [EPR4055] (ab108339)
      All lanes : Anti-CD168 antibody [EPR4055] (ab108339) at 1/1000 dilution

      Lane 1 : T47-D cell lysate
      Lane 2 : MCF-7 cell lysate
      Lane 3 : SKBR-3 cell lysate
      Lane 4 : LnCaP cell lysate

      Lysates/proteins at 10 µg per lane.

      Predicted band size: 84 kDa

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD168 antibody [EPR4055] (ab108339)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD168 antibody [EPR4055] (ab108339)

      ab108339, at 1/100, staining CD168 in paraffin-embedded human testis tissue by Immunohistochemistry.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD168 antibody [EPR4055] (ab108339)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD168 antibody [EPR4055] (ab108339)

      ab108339 showing positive staining in Normal thymus tissue.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD168 antibody [EPR4055] (ab108339)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD168 antibody [EPR4055] (ab108339)

      ab108339 showing positive staining in Normal stomach tissue.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD168 antibody [EPR4055] (ab108339)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD168 antibody [EPR4055] (ab108339)

      ab108339 showing negative staining in Normal brain tissue.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD168 antibody [EPR4055] (ab108339)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD168 antibody [EPR4055] (ab108339)

      ab108339 showing positive staining in Colonic adenocarcinoma tissue.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD168 antibody [EPR4055] (ab108339)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD168 antibody [EPR4055] (ab108339)

      ab108339 showing negative staining in Normal liver tissue.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD168 antibody [EPR4055] (ab108339)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD168 antibody [EPR4055] (ab108339)

      ab108339 showing positive staining in Normal tonsil tissue.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • Anti-CD168 antibody [EPR4055] (ab108339)
      Anti-CD168 antibody [EPR4055] (ab108339)

    Protocols

    • Western blot protocols
    • Immunoprecipitation protocols
    • Immunohistochemistry protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (4)

    Publishing research using ab108339? Please let us know so that we can cite the reference in this datasheet.

    ab108339 has been referenced in 4 publications.

    • Chen YT  et al. Immunohistochemical analysis of RHAMM expression in normal and neoplastic human tissues: a cell cycle protein with distinctive expression in mitotic cells and testicular germ cells. Oncotarget 9:20941-20952 (2018). PubMed: 29765511
    • Wang K & Zhang T Prognostic significance of CD168 overexpression in colorectal cancer. Oncol Lett 12:2555-2559 (2016). IHC ; Human . PubMed: 27698827
    • Wang D  et al. Expression of the receptor for hyaluronic acid mediated motility (RHAMM) is associated with poor prognosis and metastasis in non-small cell lung carcinoma. Oncotarget 7:39957-39969 (2016). WB, IHC . PubMed: 27220886
    • Picaud L  et al. Evaluation of the effects of hyaluronic acid-carboxymethyl cellulose barrier on ovarian tumor progression. J Ovarian Res 7:40 (2014). WB ; Human . PubMed: 24739440

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-6 of 6 Abreviews or Q&A

    Question

    Could you please provide me with the concentrations for items ab124729 (lot GR82264-8) and ab108339?

    Read More

    Abcam community

    Verified customer

    Asked on Dec 03 2013

    Answer

    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.

    Read More

    Elena Valued Customer

    Abcam Scientific Support

    Answered on Dec 03 2013

    Question

    The example on the Abcam website did say that fixation with paraformaldehyde was used but the cells were NOT permeabilized. So does this mean that the antibody recognizes denatured and not native protein or is the protein on the cell surface accessible only after the fixation process ?

    Read More

    Abcam community

    Verified customer

    Asked on Jan 15 2013

    Answer

    There should not be concerns about permeabilizing cells because as soon as they are fixed with parafomraldehyde, they are essentially permeabilized. The CD168 antigen is a non-integral cell surface hyaluronan receptor that is readily accessable to the antibody.

    If you have not already done so, you might try an overnight incubation at 4 degrees C with the primary.

    Read More

    Abcam Scientific Support

    Answered on Jan 15 2013

    Question

    Rabbit Mab, ab 108339 (clone EPR4055) – Do you have a protocol for using this antibody for flow cytometry ? I have tried this antibody using standard staining protocol and do not see any staining. Do the cells need to be fixed first (see 2D6 antibody) ?

    Read More

    Abcam community

    Verified customer

    Asked on Jan 14 2013

    Answer

    A link below is our Flow Cytometry protocol, which we use routinely for in-house testing.
    http://www.epitomics.com/products/support/fc
    Our protocol is for intracellular staining and we always fix the cells with 2% paraformaldehyde.

    Read More

    Abcam Scientific Support

    Answered on Jan 14 2013

    Question

    Thank you for the information.




    It is really important forthe customerto know exactly which of the four known isoforms of RHAMM ab108339 recognizes.

    In the datasheet of the other antibody (ab67003) it says that it recognizes all 4 epitopes.



    The 4 accession numbers of RHAMM isoforms are as follows:

    a) NM_001142556 NP_001136028.1

    b) NM_012484 NP_036616

    c) NM_012485 NP_0.6617

    d) NM_001142557 NP_001136029


    Kindly advise.



    Thanks in advance for your assistance and reply.



    Looking forward to meeting you next week (during the distributor meeting).





    Kind Regards,

    Read More

    Abcam community

    Verified customer

    Asked on Sep 03 2012

    Answer

    Thank you for your email.
    I can confirm that ab108339 (Anti-CD168 antibody) would recognize all 4 isoforms of CD168/RHAMM.
    If you need any further assistance in the future, please do not hesitate to contact me.

    Read More

    Abcam Scientific Support

    Answered on Sep 03 2012

    Question

    Please send IHC-P protocol

    Read More

    Abcam community

    Verified customer

    Asked on Jul 23 2012

    Answer

    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.

    Read More

    Abcam Scientific Support

    Answered on Jul 23 2012

    Question

    Customer kindly asked
    - the immunogenic info of antibody.
    - Whether cells needs to permeabilized and fixed in FACS

    Read More

    Abcam community

    Verified customer

    Asked on Jan 24 2012

    Answer

    Thank you for contacting us.
    The target CD168 is a cell surface protein. There no need for cell permeabilization and fixation (do not wait longer than an hour). Cell fixatiion is advised if there is a wait longer than an hour between analysis. Avoid the fixation if cells need to be viable.
    Please click to link to find more information;
    https://www.abcam.com/index.html?pageconfig=resource&rid=11380
    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Read More

    Abcam Scientific Support

    Answered on Jan 24 2012

    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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