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    glypican-3-antibody-sp86-ab95363.pdf

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Stem Cells Lineage Markers Mesoderm
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Glypican 3 antibody [SP86] (ab95363)

  • Datasheet
  • SDS
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Western blot - Anti-Glypican 3 antibody [SP86] (ab95363)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glypican 3 antibody [SP86] (ab95363)
  • Immunocytochemistry/ Immunofluorescence - Anti-Glypican 3 antibody [SP86] (ab95363)
  • Flow Cytometry - Anti-Glypican 3 antibody [SP86] (ab95363)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glypican 3 antibody [SP86] (ab95363)
  • Flow Cytometry - Anti-Glypican 3 antibody [SP86] (ab95363)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glypican 3 antibody [SP86] (ab95363)
  • Immunocytochemistry/ Immunofluorescence - Anti-Glypican 3 antibody [SP86] (ab95363)
  • Anti-Glypican 3 antibody [SP86] (ab95363)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [SP86] to Glypican 3
  • Suitable for: ICC/IF, Flow Cyt, WB, IHC-P
  • Knockout validated
  • Reacts with: Human

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Overview

  • Product name

    Anti-Glypican 3 antibody [SP86]
    See all Glypican 3 primary antibodies
  • Description

    Rabbit monoclonal [SP86] to Glypican 3
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, Flow Cyt, WB, IHC-Pmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human Glypican 3 aa 500 to the C-terminus (C terminal). The exact sequence is proprietary.
    Database link: P51654

  • Positive control

    • Human liver cancer tissueThis antibody gave a positive result when used in the following formaldehyde fixed cell lines: HepG2. ICC/IF: HepG2Flow Cyt: HepG2
  • 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.

    This product is FOR RESEARCH USE ONLY. For commercial use, please contact partnerships@abcam.com.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage buffer

    pH: 7.20
    Preservative: 0.1% Sodium azide
    Constituents: 1% BSA, PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    SP86
  • Isotype

    IgG
  • Research areas

    • Stem Cells
    • Lineage Markers
    • Mesoderm
    • Signal Transduction
    • Cytoskeleton / ECM
    • HSPGs
    • Cancer
    • Tumor biomarkers
    • Other
    • Developmental Biology
    • Lineage specification
    • Mesoderm

Associated products

  • Alternative Versions

    • Anti-Glypican 3 antibody [SP86] - BSA and Azide free (ab238804)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

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

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab95363 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
1/200.
Flow Cyt
1/80.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

WB
Use at an assay dependent concentration. Predicted molecular weight: 66 kDa.
IHC-P
1/100. Perform heat mediated antigen retrieval (boil tissue section in 10mM citrate buffer, pH 6.0 for 10 min followed by cooling at RT for 20 min).
Notes
ICC/IF
1/200.
Flow Cyt
1/80.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

WB
Use at an assay dependent concentration. Predicted molecular weight: 66 kDa.
IHC-P
1/100. Perform heat mediated antigen retrieval (boil tissue section in 10mM citrate buffer, pH 6.0 for 10 min followed by cooling at RT for 20 min).

Target

  • Function

    Cell surface proteoglycan that bears heparan sulfate. Inhibits the dipeptidyl peptidase activity of DPP4. May be involved in the suppression/modulation of growth in the predominantly mesodermal tissues and organs. May play a role in the modulation of IGF2 interactions with its receptor and thereby modulate its function. May regulate growth and tumor predisposition.
  • Tissue specificity

    Highly expressed in lung, liver and kidney.
  • Involvement in disease

    Defects in GPC3 are the cause of Simpson-Golabi-Behmel syndrome type 1 (SGBS1) [MIM:312870]; also known as Simpson dysmorphia syndrome (SDYS). SGBS is a condition characterized by pre- and postnatal overgrowth (gigantism) with visceral and skeletal anomalies.
  • Sequence similarities

    Belongs to the glypican family.
  • Cellular localization

    Cell membrane and Secreted > extracellular space.
  • Target information above from: UniProt accession P51654 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 2719 Human
    • Omim: 300037 Human
    • SwissProt: P51654 Human
    • Unigene: 644108 Human
    • Alternative names

      • DGSX antibody
      • Glypican proteoglycan 3 antibody
      • Glypican-3 [Precursor] antibody
      • Gpc3 antibody
      • GPC3_HUMAN antibody
      • GTR2 2 antibody
      • GTR2-2 antibody
      • Heparan sulphate proteoglycan antibody
      • Intestinal protein OCI 5 antibody
      • Intestinal protein OCI-5 antibody
      • MXR7 antibody
      • OCI 5 antibody
      • OCI-5 antibody
      • OCI5 antibody
      • SDYS antibody
      • Secreted glypican-3 antibody
      • SGB antibody
      • SGBS antibody
      • SGBS1 antibody
      see all

    Images

    • Western blot - Anti-Glypican 3 antibody [SP86] (ab95363)
      Western blot - Anti-Glypican 3 antibody [SP86] (ab95363)

      Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
      Lane 2: GPC3 knockout HAP1 whole cell lysate (20 µg)
      Lane 3: HepG2 whole cell lysate (20 µg)

      Lanes 1 - 3: Merged signal (red and green). Green - ab95363 observed at 70 kDa. Red - loading control, ab130007, observed at 125kDa.


      ab95363 was shown to specifically react with Glypican 3 in wild-type HAP1 cells as signal was lost in GPC3 knockout cells. Wild-type and GPC3 knockout samples were subjected to SDS-PAGE. ab95363 and ab130007 (Mouse anti-vinculin loading control) were incubated overnight at 4°C at 1/1000 and 1/20000 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 1/10000 dilution for 1 hour at room temperature before imaging.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glypican 3 antibody [SP86] (ab95363)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glypican 3 antibody [SP86] (ab95363)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human hepatocellular carcinoma tissue sections labeling Glypican 3 with ab95363 at 1:100 dilution (4.66 ?g/ml). Heat mediated antigen retrieval with sodium citrate buffer (pH 6.0, epitope retrieval solution 1) for 10mins. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Hematoxylin was used as a counterstain. Positive staining on human hepatocellular carcinoma, performed on a Leica Biosystems BONDTM RX instrument.
      The section was incubated with ab95363 for 30 mins at room temperature.

    • Immunocytochemistry/ Immunofluorescence - Anti-Glypican 3 antibody [SP86] (ab95363)
      Immunocytochemistry/ Immunofluorescence - Anti-Glypican 3 antibody [SP86] (ab95363)

      Immunocytochemistry/ Immunofluorescence analysis of HepG2 (human hepatocellular carcinoma epithelial cell) cells labeling Glypican 3 with purified ab95363 at 1/200 (2.3 µg/ml). Cells were fixed in 100% Methanol and permeabilized with None. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/ml) dilution. DAPI was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

    • Flow Cytometry - Anti-Glypican 3 antibody [SP86] (ab95363)
      Flow Cytometry - Anti-Glypican 3 antibody [SP86] (ab95363)

      Flow cytometry analysis of HepG2 (human hepatocellular carcinoma) labeling Glypican 3 with purified ab95363 at 1/80 dilution (5.825 µg/ml) (red). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as a secondary antibody. Isotype control - Rabbit monoclonal IgG (ab172730) (Black). Unlableled control -Unlabelled cells (blue).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glypican 3 antibody [SP86] (ab95363)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glypican 3 antibody [SP86] (ab95363)

      Immunohistochemical staining of human liver hepatocellular carcinoma with ab95363.

    • Flow Cytometry - Anti-Glypican 3 antibody [SP86] (ab95363)
      Flow Cytometry - Anti-Glypican 3 antibody [SP86] (ab95363)

      Flow cytometric analysis of rabbit anti-Glypican 3 (SP86) antibody ab98363 (1/100) in HEPG2 cellls (green) compared to negative control of rabbit IgG (blue).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glypican 3 antibody [SP86] (ab95363)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glypican 3 antibody [SP86] (ab95363)
      ab95363, at 1/100 dilution, staining Glypican 3 in formalin-fixed, paraffin-embedded Human liver cancer tissue by Immunohistochemistry.
    • Immunocytochemistry/ Immunofluorescence - Anti-Glypican 3 antibody [SP86] (ab95363)
      Immunocytochemistry/ Immunofluorescence - Anti-Glypican 3 antibody [SP86] (ab95363)

      ICC/IF image of ab95363 stained HepG2 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab95363 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

    • Anti-Glypican 3 antibody [SP86] (ab95363)
      Anti-Glypican 3 antibody [SP86] (ab95363)

    Protocols

    • Flow cytometry protocols
    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (2)

    Publishing research using ab95363? Please let us know so that we can cite the reference in this datasheet.

    ab95363 has been referenced in 2 publications.

    • Dong W  et al. Prognostic Nomogram for Sorafenib Benefit in Hepatitis B Virus-Related Hepatocellular Carcinoma After Partial Hepatectomy. Front Oncol 10:605057 (2020). PubMed: 33643907
    • Wang Y  et al. Transcobalamin I: a novel prognostic biomarker of neoadjuvant chemotherapy in locally advanced hypopharyngeal squamous cell cancers. Onco Targets Ther 11:4253-4261 (2018). PubMed: 30100732

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