Recombinant
RabMAb

Recombinant Anti-CD44 antibody [EPR18668] (ab189524)

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Overview

  • Product name

    Anti-CD44 antibody [EPR18668]
    See all CD44 primary antibodies

  • Description

    Rabbit monoclonal [EPR18668] to CD44

  • Host species

    Rabbit

  • Tested applications

    Suitable for: ICC/IF, Flow Cyt, WB, IHC-P, IPmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Recombinant fragment within Human CD44 aa 1-250. The exact sequence is proprietary.
    Database link: P16070

  • Positive control

    • WB: Human fetal brain, fetal heart, fetal kidney and fetal spleen lysates; Human thymus and skin lysates; A549, U-87 MG, C6, RAW 264.7, PC-12 and NIH/3T3 whole cell lysates; Mouse brain and spleen lysates; Rat brain, heart, kidney and spleen lysates; IHC-P: Human breast, kidney, tonsil and breast cancer tissues; Mouse colon, stomach and spleen tissues; Rat stomach and spleen tissues; IP: A549 whole cell lysate. ICC/IF: MDA-MB-231 cell line. Flow Cytometry: MDA-MB-231 cell line.

  • 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.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab189524 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
ICC/IF 1/250.
Flow Cyt 1/40.
WB 1/1000. Detects a band of approximately 82 kDa (predicted molecular weight: 82 kDa).
IHC-P 1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IP 1/30.

Target

  • Function

    Receptor for hyaluronic acid (HA). Mediates cell-cell and cell-matrix interactions through its affinity for HA, and possibly also through its affinity for other ligands such as osteopontin, collagens, and matrix metalloproteinases (MMPs). Adhesion with HA plays an important role in cell migration, tumor growth and progression. Also involved in lymphocyte activation, recirculation and homing, and in hematopoiesis. Altered expression or dysfunction causes numerous pathogenic phenotypes. Great protein heterogeneity due to numerous alternative splicing and post-translational modification events.

  • Tissue specificity

    Isoform 10 (epithelial isoform) is expressed by cells of epithelium and highly expressed by carcinomas. Expression is repressed in neuroblastoma cells.

  • Sequence similarities

    Contains 1 Link domain.

  • Domain

    The lectin-like LINK domain is responsible for hyaluronan binding.

  • Post-translational
    modifications

    Proteolytically cleaved in the extracellular matrix by specific proteinases (possibly MMPs) in several cell lines and tumors.
    N-glycosylated.
    O-glycosylated; contains more-or-less-sulfated chondroitin sulfate glycans, whose number may affect the accessibility of specific proteinases to their cleavage site(s).
    Phosphorylated; activation of PKC results in the dephosphorylation of Ser-706 (constitutive phosphorylation site), and the phosphorylation of Ser-672.

  • Cellular localization

    Membrane.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • LHR antibody
    • BA-1 antibody
    • CD 44 antibody
    • CD44 antibody
    • CD44 antigen antibody
    • CD44 molecule (Indian blood group) antibody
    • CD44 molecule antibody
    • CD44_HUMAN antibody
    • CDw44 antibody
    • CDW44 antigen antibody
    • Cell surface glycoprotein CD44 antibody
    • chondroitin sulfate proteoglycan 8 antibody
    • CSPG8 antibody
    • ECMR-III antibody
    • Epican antibody
    • Extracellular matrix receptor III antibody
    • GP90 lymphocyte homing/adhesion receptor antibody
    • HCELL antibody
    • hematopoietic cell E- and L-selectin ligand antibody
    • Heparan sulfate proteoglycan antibody
    • Hermes antigen antibody
    • homing function and Indian blood group system antibody
    • HSA antibody
    • HUTCH-I antibody
    • HUTCH1 antibody
    • HUTCHI antibody
    • Hyaluronate receptor antibody
    • IN antibody
    • INLU-related p80 Glycoprotein antibody
    • MC56 antibody
    • MDU2 antibody
    • MDU3 antibody
    • MGC10468 antibody
    • MIC4 antibody
    • MUTCH I antibody
    • MUTCH1 antibody
    • PGP-1 antibody
    • PGP-I antibody
    • PGP1 antibody
    • Phagocytic glycoprotein 1 antibody
    • Phagocytic glycoprotein I antibody
    • Soluble CD44 antibody
    see all

Images

  • Immunocytochemistry/ Immunofluorescence - Anti-CD44 antibody [EPR18668] (ab189524)
    Immunocytochemistry/ Immunofluorescence - Anti-CD44 antibody [EPR18668] (ab189524)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized MDA-MB-231 (Human breast adenocarcinoma cell line) labeling CD44 with ab189524 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal image showing membranous staining on MDA-MB-231 cell line.

    The nuclear counterstain is DAPI (blue).

    Tubulin is detected with Mouse monoclonal [DM1A] to alpha Tubulin - Microtubule Marker (Alexa Fluor® 594) at 1/200 dilution.

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD44 antibody [EPR18668] (ab189524)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD44 antibody [EPR18668] (ab189524)

    Immunohistochemical analysis of paraffin-embedded human breast tissue labeling CD44 with ab189524 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Membranous and weak cytoplasmic staining on human breast [PMID: 20103682].

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD44 antibody [EPR18668] (ab189524)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD44 antibody [EPR18668] (ab189524)

    Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling CD44 with ab189524 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Membranous and weak cytoplasmic staining on human kidney.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

  • Western blot - Anti-CD44 antibody [EPR18668] (ab189524)
    Western blot - Anti-CD44 antibody [EPR18668] (ab189524)
    All lanes : Anti-CD44 antibody [EPR18668] (ab189524) at 1/1000 dilution

    Lane 1 : Human fetal brain lysate
    Lane 2 : Human fetal heart lysate
    Lane 3 : Human fetal kidney lysate
    Lane 4 : Human fetal spleen lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/10000 dilution

    Predicted band size: 82 kDa
    Observed band size: 82 kDa


    Exposure time: 5 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

     

  • Western blot - Anti-CD44 antibody [EPR18668] (ab189524)
    Western blot - Anti-CD44 antibody [EPR18668] (ab189524)
    All lanes : Anti-CD44 antibody [EPR18668] (ab189524) at 1/2000 dilution

    Lane 1 : Human thymus lysate
    Lane 2 : Human skin lysate
    Lane 3 : A549 (Human lung carcinoma cell line) whole cell lysate
    Lane 4 : U-87 MG (Human glioblastoma-astrocytoma epithelial cell line) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Lanes 1-2 : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/10000 dilution
    Lanes 3-4 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 82 kDa
    Observed band size: 82 kDa


    Exposure time: 1 second


    Blocking/Dilution buffer: 5% NFDM/TBST.

    Due to high sequence homology between CD44 isoforms, within the immunogen region, this antibody cross reacts with most isoforms of CD44. The staining pattern is consistent with reference (PMID 19507218).

  • Western blot - Anti-CD44 antibody [EPR18668] (ab189524)
    Western blot - Anti-CD44 antibody [EPR18668] (ab189524)
    Lanes 1-6 : Anti-CD44 antibody [EPR18668] (ab189524) at 1/1000 dilution
    Lanes 7-10 : Anti-CD44 antibody [EPR18668] (ab189524) at 1/20000 dilution

    Lane 1 : Mouse brain lysate at 10 µg
    Lane 2 : Mouse spleen lysate at 10 µg
    Lane 3 : Rat brain lysate at 10 µg
    Lane 4 : Rat heart lysate at 10 µg
    Lane 5 : Rat kidney lysate at 10 µg
    Lane 6 : Rat spleen lysate at 10 µg
    Lane 7 : C6 (Rat glial tumor cell line) whole cell lysate
    Lane 8 : RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
    Lane 9 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
    Lane 10 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate

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

    Predicted band size: 82 kDa
    Observed band size: 82 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1, 2, 3, 4, 5 and 6: 5 seconds; Lane 7, 8, 9 and 10: 1 second.

  • Flow Cytometry - Anti-CD44 antibody [EPR18668] (ab189524)
    Flow Cytometry - Anti-CD44 antibody [EPR18668] (ab189524)

    Flow cytometric analysis of 4% paraformaldehyde-fixed MDA-MB-231 (Human breast adenocarcinoma cell line) cell line labeling CD44 with ab189524 at 1/40 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody.

     

  • Immunoprecipitation - Anti-CD44 antibody [EPR18668] (ab189524)
    Immunoprecipitation - Anti-CD44 antibody [EPR18668] (ab189524)

    CD44v6 was immunoprecipitated from 1 mg of A549 (Human lung carcinoma cell line) whole cell lysate with ab189524 at 1/25 dilution. Western blot was performed from the immunoprecipitate using ab189524 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: A549 whole cell lysate 10 μg (Input).

    Lane 2: ab189524 IP in A549 whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab189524 in A549 whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 1 second.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD44 antibody [EPR18668] (ab189524)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD44 antibody [EPR18668] (ab189524)

    Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling CD44 with ab189524 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Membranous and weak cytoplasmic staining on human tonsil.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD44 antibody [EPR18668] (ab189524)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD44 antibody [EPR18668] (ab189524)

    Immunohistochemical analysis of paraffin-embedded human breast cancer tissue labeling CD44 with ab189524 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Membranous and weak cytoplasmic staining on human breast cancer [PMID: 15867228].

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD44 antibody [EPR18668] (ab189524)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD44 antibody [EPR18668] (ab189524)

    Immunohistochemical analysis of paraffin-embedded mouse colon tissue labeling CD44 with ab189524 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Membranous and weak cytoplasmic staining on mouse colon.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD44 antibody [EPR18668] (ab189524)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD44 antibody [EPR18668] (ab189524)

    Immunohistochemical analysis of paraffin-embedded mouse stomach tissue labeling CD44 with ab189524 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Membranous and weak cytoplasmic staining on mouse stomach.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD44 antibody [EPR18668] (ab189524)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD44 antibody [EPR18668] (ab189524)

    Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling CD44 with ab189524 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Membranous and weak cytoplasmic staining on mouse spleen.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD44 antibody [EPR18668] (ab189524)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD44 antibody [EPR18668] (ab189524)

    Immunohistochemical analysis of paraffin-embedded rat stomach tissue labeling CD44 with ab189524 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Membranous staining on rat stomach.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD44 antibody [EPR18668] (ab189524)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD44 antibody [EPR18668] (ab189524)

    Immunohistochemical analysis of paraffin-embedded rat spleen tissue labeling CD44 with ab189524 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Membranous and weak cytoplasmic staining on rat spleen.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

References

This product has been referenced in:

  • Zhang C & Wang Y Metformin attenuates cells stemness and epithelial-mesenchymal transition in colorectal cancer cells by inhibiting the Wnt3a/ß-catenin pathway. Mol Med Rep 19:1203-1209 (2019). Read more (PubMed: 30569135) »
  • Zhou ZQ  et al. Adipose extracellular matrix promotes skin wound healing by inducing the differentiation of adipose-derived stem cells into fibroblasts. Int J Mol Med 43:890-900 (2019). Read more (PubMed: 30535488) »
See all 3 Publications for this product

Publishing research using ab189524? Please let us know so that we can cite the reference in this datasheet.

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