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  1. Link

    cd44-antibody-blr038f-ab243894.pdf

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Immunology Cell Type Markers CD Adhesion
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Validated using a knockout cell lineRecombinant

Recombinant Anti-CD44 antibody [BLR038F] (ab243894)

  • Datasheet
  • SDS
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD44 antibody [BLR038F] (ab243894)
  • Western blot - Anti-CD44 antibody [BLR038F] (ab243894)
  • Immunoprecipitation - Anti-CD44 antibody [BLR038F] (ab243894)
  • Flow Cytometry - Anti-CD44 antibody [BLR038F] (ab243894)
  • Immunocytochemistry - Anti-CD44 antibody [BLR038F] (ab243894)
  • Western blot - Anti-CD44 antibody [BLR038F] (ab243894)
  • Western blot - Anti-CD44 antibody [BLR038F] (ab243894)
  • Anti-CD44 antibody [BLR038F] (ab243894)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [BLR038F] to CD44
  • Suitable for: Flow Cyt, ICC, IHC-P, IP, WB
  • Knockout validated
  • Reacts with: Mouse, Human

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Overview

  • Product name

    Anti-CD44 antibody [BLR038F]
    See all CD44 primary antibodies
  • Description

    Rabbit monoclonal [BLR038F] to CD44
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Flow Cyt, ICC, IHC-P, IP, WBmore details
  • Species reactivity

    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide within Human CD44 aa 700 to the C-terminus (C terminal). The exact immunogen sequence used to generate this antibody is proprietary information. If additional detail on the immunogen is needed to determine the suitability of the antibody for your needs, please contact our Scientific Support team to discuss your requirements. NP_000601.3 and Gene ID 960.
    Database link: P16070-1

  • Positive control

    • WB: 786-O, KG-1, HeLa and A549 cell lysates. EL4 and BW5147.3 cell lysates. IHC-P: Human head and neck squamous cell carcinoma tissue. ICC: HCT116 cells. Flow Cyt: HeLa cells.
  • General notes

    This product is sold under License from Bethyl Laboratories, Inc.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.8
    Preservative: 0.09% Sodium azide
    Constituents: 98% Borate buffered saline, 0.1% BSA
  • Concentration information loading...
  • Purification notes

    Recombinant antibody was purified from cell culture supernatant.
  • Clonality

    Monoclonal
  • Clone number

    BLR038F
  • Isotype

    IgG
  • Research areas

    • Immunology
    • Cell Type Markers
    • CD
    • Adhesion
    • Stem Cells
    • Mesenchymal Stem Cells
    • Surface Molecules
    • Cancer
    • Tumor immunology
    • CD markers
    • Cancer
    • Tumor biomarkers
    • Other
    • Stem Cells
    • Hematopoietic Progenitors
    • Lymphoid
    • T Lymphocytic Lineage
    • Stem Cells
    • Hematopoietic Progenitors
    • Myeloid
    • Dendritic Cell Lineage
    • Stem Cells
    • Hematopoietic Progenitors
    • Myeloid
    • Neutrophil Lineage
    • Kits/ Lysates/ Other
    • Kits
    • ELISA Kits
    • ELISA Kits
    • CD markers ELISA kits

Associated products

  • KO cell lines

    • Human CD44 knockout HeLa cell line (ab262515)
  • KO cell lysates

    • Human CD44 knockout HeLa cell lysate (ab263938)
  • Recombinant Protein

    • Recombinant human CD44 protein (Active) (ab173996)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab243894 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
Flow Cyt
Use 1µl for 106 cells.
ICC
1/100 - 1/500.
IHC-P
1/100 - 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
IP
Use at an assay dependent concentration.

Use 20µl/mg lysate.

WB
1/1000.
Notes
Flow Cyt
Use 1µl for 106 cells.
ICC
1/100 - 1/500.
IHC-P
1/100 - 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
IP
Use at an assay dependent concentration.

Use 20µl/mg lysate.

WB
1/1000.

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.
  • Target information above from: UniProt accession P16070 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 960 Human
    • Entrez Gene: 12505 Mouse
    • Omim: 107269 Human
    • SwissProt: P16070 Human
    • SwissProt: P15379 Mouse
    • Unigene: 502328 Human
    • Unigene: 423621 Mouse
    • 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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD44 antibody [BLR038F] (ab243894)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD44 antibody [BLR038F] (ab243894)
      Formalin-fixed, paraffin-embedded human head and neck squamous cell carcinoma tissue stained for CD44 using ab243894 at 1/500 dilution in immunohistochemical analysis. A HRP-conjugated goat anti-rabbit antibody was used as the secondary. DAB staining.
    • Western blot - Anti-CD44 antibody [BLR038F] (ab243894)
      Western blot - Anti-CD44 antibody [BLR038F] (ab243894)
      All lanes : Anti-CD44 antibody [BLR038F] (ab243894) at 1/1000 dilution

      Lane 1 : Wild-type HeLa cell lysate
      Lane 2 : Knockout CD44 HeLa cell lysate
      Lane 3 : A549 cell lysate
      Lane 4 : LnCAP cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Observed band size: 80 kDa why is the actual band size different from the predicted?



      Lanes 1 - 4: Merged signal (red and green). Green - ab243894 observed at 80 kDa. Red - loading control, ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.

      ab243894 was shown to react with CD44 in wild-type HeLa cells in western blot. Loss of signal was observed when CD44 knockout sample was used. Wild-type HeLa and CD44 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab243894 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4°C at a 1 in 1000 Dilution and a 1 in 20000 dilution respectively. Blots were incubated 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 in 20000 dilution for 1 hour at room temperature before imaging.

    • Immunoprecipitation - Anti-CD44 antibody [BLR038F] (ab243894)
      Immunoprecipitation - Anti-CD44 antibody [BLR038F] (ab243894)
      CD44 was immunoprecipitated from 1.0 mg HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab243894 at 20 µl per reaction. Western blot was performed on the immunoprecipitate using ab243894 at 1/1000 dilution.

      Lane 1: ab243894 IP in HeLa whole cell lysate.

      Lane 2: Control IgG in HeLa whole cell lysate.

      Detection: Chemiluminescence with an exposure time of 3 seconds.

    • Flow Cytometry - Anti-CD44 antibody [BLR038F] (ab243894)
      Flow Cytometry - Anti-CD44 antibody [BLR038F] (ab243894)
      Flow Cytometry analysis of HeLa cells labeling CD44 with ab243894 (red). Cells were fixed in 4% formaldehyde and permeabilized with 90% methanol. Isotype control (unshaded). Secondary: DyLight® 650-conjugated goat anti-rabbit IgG.
    • Immunocytochemistry - Anti-CD44 antibody [BLR038F] (ab243894)
      Immunocytochemistry - Anti-CD44 antibody [BLR038F] (ab243894)
      Formalin-fixed, paraffin-embedded HCT 116 (human colorectal carcinoma cell line) cells labeling CD44 using ab243894 at 1/500 dilution in ICC analysis. A HRP-conjugated goat-anti rabbit IgG was used as the secondary. DAB staining.
    • Western blot - Anti-CD44 antibody [BLR038F] (ab243894)
      Western blot - Anti-CD44 antibody [BLR038F] (ab243894)
      All lanes : Anti-CD44 antibody [BLR038F] (ab243894) at 1/1000 dilution

      Lane 1 : HeLa whole cell lysate
      Lane 2 : 786-O whole cell lysate
      Lane 3 : HEK-293T whole cell lysate
      Lane 4 : Jurkat-3 whole cell lysate
      Lane 5 : A549 whole cell lysate
      Lane 6 : MCF7 whole cell lysate
      Lane 7 : HepG2 whole cell lysate
      Lane 8 : KG-1 whole cell lysate
      Lane 9 : RPMI 8226 whole cell lysate
      Lane 10 : MOLT-4 whole cell lysate

      Lysates/proteins at 50 µg per lane.

      Secondary
      All lanes : HRP-conjugated goat anti-rabbit IgG antibody


      Detection: chemiluminescence with an exposure time of 3 seconds.

    • Western blot - Anti-CD44 antibody [BLR038F] (ab243894)
      Western blot - Anti-CD44 antibody [BLR038F] (ab243894)
      All lanes : Anti-CD44 antibody [BLR038F] (ab243894) at 1/1000 dilution

      Lane 1 : BW5147.3 cell lysate
      Lane 2 : EL4 cell lysate

      Lysates/proteins at 50 µg per lane.

      Secondary
      All lanes : HRP-conjugated goat anti-rabbit IgG

      Exposure time: 10 seconds


      Detection method: Chemiluminescence

    • Anti-CD44 antibody [BLR038F] (ab243894)
      Anti-CD44 antibody [BLR038F] (ab243894)

    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

    • SDS download

    • Datasheet download

      Download

    References (1)

    Publishing research using ab243894? Please let us know so that we can cite the reference in this datasheet.

    ab243894 has been referenced in 1 publication.

    • Li S  et al. Schisandrin B inhibits epithelial-mesenchymal transition and stemness of large-cell lung cancer cells and tumorigenesis in xenografts via inhibiting the NF-?B and p38 MAPK signaling pathways. Oncol Rep 45:N/A (2021). PubMed: 33907830

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