Recombinant
RabMAb

Recombinant Anti-Podoplanin / gp36 antibody [EPR22182] (ab236529)

Overview

  • Product name

    Anti-Podoplanin / gp36 antibody [EPR22182]
    See all Podoplanin / gp36 primary antibodies
  • Description

    Rabbit monoclonal [EPR22182] to Podoplanin / gp36
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, Flow Cyt, IPmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Recombinant fragment within Human Podoplanin/ gp36 aa 1-150. The exact sequence is proprietary.
    Database link: Q86YL7

  • Positive control

    • WB: U-2 OS and U-87 MG whole cell lysates; Human seminoma, tonsil, lymph node and stomach lysates. IHC-P: Human colon, mesothelioma and Kaposi’s sarcoma tissues. Flow Cyt: U-87 MG and U-2 OS cells. IP: U-2 OS whole cell lysate.
  • 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 ab236529 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. Detects a band of approximately 38 kDa (predicted molecular weight: 16 kDa).
IHC-P 1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Flow Cyt 1/300.
IP 1/30.

Target

  • Function

    May be involved in cell migration and/or actin cytoskeleton organization. When expressed in keratinocytes, induces changes in cell morphology with transfected cells showing an elongated shape, numerous membrane protrusions, major reorganization of the actin cytoskeleton, increased motility and decreased cell adhesion. Required for normal lung cell proliferation and alveolus formation at birth. Induces platelet aggregation. Does not have any effect on folic acid or amino acid transport. Does not function as a water channel or as a regulator of aquaporin-type water channels.
  • Tissue specificity

    Highly expressed in placenta, lung, skeletal muscle and brain. Weakly expressed in brain, kidney and liver. In placenta, expressed on the apical plasma membrane of endothelium. In lung, expressed in alveolar epithelium. Up-regulated in colorectal tumors and expressed in 25% of early oral squamous cell carcinomas.
  • Sequence similarities

    Belongs to the podoplanin family.
  • Post-translational
    modifications

    Extensively O-glycosylated. Contains sialic acid residues. O-glycosylation is necessary for platelet aggregation activity.
    The N-terminus is blocked.
  • Cellular localization

    Membrane. Cell projection > filopodium membrane. Cell projection > lamellipodium membrane. Cell projection > microvillus membrane. Cell projection > ruffle membrane. Localized to actin-rich microvilli and plasma membrane projections such as filopodia, lamellipodia and ruffles.
  • Information by UniProt
  • Database links

  • Alternative names

    • Aggrus antibody
    • Glycoprotein 36 KD antibody
    • Glycoprotein 36 antibody
    • gp 36 antibody
    • GP 38 antibody
    • GP 40 antibody
    • gp36 antibody
    • GP38 antibody
    • GP40 antibody
    • HT1A 1 antibody
    • HT1A1 antibody
    • hT1alpha 1 antibody
    • hT1alpha 2 antibody
    • hT1alpha1 antibody
    • hT1alpha2 antibody
    • Lung type I cell membrane associated glycoprotein antibody
    • Lung type I cell membrane associated glycoprotein isoform a antibody
    • Lung type I cell membrane associated glycoprotein T1A 2 antibody
    • OTS 8 antibody
    • OTS8 antibody
    • OTTHUMP00000009640 antibody
    • OTTHUMP00000044504 antibody
    • PA2.26 antibody
    • PA2.26 antigen antibody
    • Pdpn antibody
    • PDPN_HUMAN antibody
    • Podoplanin antibody
    • PSEC0003 antibody
    • PSEC0025 antibody
    • T1 alpha antibody
    • T1 ALPHA GENE antibody
    • T1-alpha antibody
    • T1A 2 antibody
    • T1A antibody
    • TI1A antibody
    • TIA 2 antibody
    • TIA2 antibody
    see all

Images

  • Immunohistochemical analysis of paraffin-embedded human colon tissue labeling Podoplanin / gp36 with ab236529 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Positive staining in lymphatic endothelial cells of human colon (PMID: 28101903; PMID: 27798887) is observed. 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.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Podoplanin / gp36 was immunoprecipitated from 0.35 mg of U-2 OS (human bone osteosarcoma epithelial cell line) whole cell lysate with ab236529 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab236529 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.

    Lane 1: U-2 OS whole cell lysate 10 µg (Input). 

    Lane 2: ab236529 IP in U-2 OS whole cell lysate. 

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab236529 in U-2 OS whole cell lysate.

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

    Exposure time: 30 seconds.

  • Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized U-2 OS (human bone osteosarcoma epithelial cell line) cell line labeling Podoplanin / gp36 with ab236529 at 1/300 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.

  • Immunohistochemical analysis of paraffin-embedded human mesothelioma tissue labeling Podoplanin / gp36 with ab236529 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Membranous and weak cytoplasmic staining in tumor cells of human mesothelioma (PMID: 16670463) is observed. 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.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • All lanes : Anti-Podoplanin / gp36 antibody [EPR22182] (ab236529) at 1/1000 dilution

    Lane 1 : Human seminoma lysate
    Lane 2 : Human tonsil lysate
    Lane 3 : Human lymph node lysate
    Lane 4 : Human stomach lysate

    Lysates/proteins at 20 µg per lane.

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

    Predicted band size: 16 kDa
    Observed band size: 38 kDa
    why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The molecular mass observed is consistent with what has been described in the literature (PMID: 10408868; PMID: 17279584).

  • Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized U-87 MG (human glioblastoma-astrocytoma epithelial cell line) cell line labeling Podoplanin / gp36 with ab236529 at 1/300 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.

  • All lanes : Anti-Podoplanin / gp36 antibody [EPR22182] (ab236529) at 1/1000 dilution

    Lane 1 : U-2 OS (human bone osteosarcoma epithelial cell line) whole cell lysate
    Lane 2 : HUVEC (human umbilical vein endothelial cell line) whole cell lysate
    Lane 3 : U-87 MG (human glioblastoma-astrocytoma epithelial cell line) whole cell lysate

    Lysates/proteins at 20 µg per lane.

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

    Predicted band size: 16 kDa
    Observed band size: 38 kDa why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The molecular mass observed is consistent with what has been described in the literature (PMID: 10408868; PMID: 17279584).

    Negative control: HUVEC (PMID: 21920451).

  • Immunohistochemical analysis of paraffin-embedded human Kaposi’s sarcoma tissue labeling Podoplanin / gp36 with ab236529 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Membranous and cytoplasmic staining in tumor cells of human Kaposi’s sarcoma (PMID: 11950918) is observed. 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.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

References

ab236529 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

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Application
Western blot
Sample
Dog Cell lysate - whole cell (Canine osteosarcoma cells)
Gel Running Conditions
Reduced Denaturing
Loading amount
37.5 µg
Specification
Canine osteosarcoma cells
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

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Submitted Nov 20 2018

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