Anti-Podoplanin / gp36 antibody [18H5] (ab10288)

Overview

  • Product nameAnti-Podoplanin / gp36 antibody [18H5]
    See all Podoplanin / gp36 primary antibodies
  • Description
    Mouse monoclonal [18H5] to Podoplanin / gp36
  • Tested applicationsIHC-P, ICC/IF, WB, Flow Cyt, IHC-Frmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Full length protein corresponding to Podoplanin/ gp36. MDCK cells stably expressing Podoplanin/ gp36 protein.
    Database link: Q86YL7

  • Positive control
    • In Western blot, this antibody gave a positive signal in human placenta and skeletal muscle tissue lysates.

Properties

Applications

Our Abpromise guarantee covers the use of ab10288 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
IHC-P Use a concentration of 6 - 30 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF Use a concentration of 1 - 2 µg/ml.
WB Use a concentration of 5 µg/ml. Detects a band of approximately 35 kDa (predicted molecular weight: 24 kDa).
Flow Cyt Use 1µl for 106 cells.

(methanol fixed cells)

IHC-Fr Use at an assay dependent concentration. PubMed: 18987360

Target

  • FunctionMay 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 specificityHighly 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 similaritiesBelongs 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 localizationMembrane. 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
    • 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
    • TIA 2 antibody
    • TIA2 antibody
    see all

Anti-Podoplanin / gp36 antibody [18H5] images

  • ab10288 staining Podoplanin / gp36 in human differentiated cardiac ventricular cells by Flow Cytometry. Cells were fixed with paraformaldehyde and permeabilized with permeabilization buffer. The sample was incubated with the primary antibody (1/300) for 12 hours at 4°C. An Alexa Fluor® 488-conjugated goat monoclonal anti-mouse IgG (1/1500) was used as the secondary antibody.
    Gating Strategy: Isotype negative control (white).

    See Abreview

  • ab10288 at a 1/500 dilution staining human dermal lymphatic endothelial by Immunocytochemistry. The cells were grown in coverslips and fixed in formalin prior to permeabilization with tritonX-100. The cells were incubated with the primary antibody at 1/500 for 1 hour and then bound antibody was detected using a biotinylated goat anti-mouse antibody. Texas red-streptavidin was used for development.

    This image is courtesy of an Abreview submitted by Maria Navarro.

    See Abreview

  • ab10288 at 1/500 dilution staining human thymus specimen tissue sections by IHC-P. The tissue was formaldehyde fixed and a heat induced epitope retrieval step was carried out. The tisse was then blocked and incubated with the antibody. An HRP conjugated rabbit antibody was used as the secondary.

    See Abreview

  • Overlay histogram showing HCT116 cells stained with ab10288 (red line). The cells were fixed with methanol (5 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab10288, 1µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a slightly decreased signal in HCT116 cells fixed with 4% paraformaldehyde (10 min) used under the same conditions.

    Please note that Abcam does not have data for use of this antibody on non-fixed cells. We welcome any customer feedback.
  • All lanes : Anti-Podoplanin / gp36 antibody [18H5] (ab10288) at 5 µg/ml

    Lane 1 : Placenta (Human) Tissue Lysate - adult normal tissue (ab29745)
    Lane 2 : Skeletal Muscle (Human) Tissue Lysate - adult normal tissue (ab29330)

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution
    developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 24 kDa
    Observed band size : 35 kDa (why is the actual band size different from the predicted?)


    Exposure time : 20 minutes
    Podoplanin protein is extensively glycosylated. This might explain the observation of a band at a higher molecular weight than predicted.


  • Predicted band size : 24 kDa

    Western blot analysis of Podoplanin expression in human lymphatic endothelial cells (LEC) - lane 1, and HUVECs - lane 2. Total lysate of both cell types were subjected to SDS-PAGE and subsequent western blot analysis with ab10288. The antibody recognizes a protein of about 36 kDa in total lysate from LECs but not from HUVEC.

  • Immunocytochemistry/Immunofluorescence analysis of human dermal lymphatic endothelial cells (HDLEC) labelling Podoplanin / gp36 with ab10288. An Alexa Fluor® 488-conjugated goat anti-mouse was used as the secondary antibody. Nuclei counterstained with DAPI.

References for Anti-Podoplanin / gp36 antibody [18H5] (ab10288)

This product has been referenced in:
  • Jakus Z  et al. Lymphatic function is required prenatally for lung inflation at birth. J Exp Med 211:815-26 (2014). Mouse . Read more (PubMed: 24733830) »
  • Hoshino A  et al. Podoplanin-positive fibroblasts enhance lung adenocarcinoma tumor formation: podoplanin in fibroblast functions for tumor progression. Cancer Res 71:4769-79 (2011). Read more (PubMed: 21610106) »

See all 10 Publications for this product

Product Wall

Application Western blot
Loading amount 30 µg
Gel Running Conditions Reduced Denaturing (10% Bis-Tris Gel)
Sample Human Cell lysate - whole cell (SW1353 human chondrosacroma cell line)
Specification SW1353 human chondrosacroma cell line
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Jul 14 2014

Application Flow Cytometry
Fixation Paraformaldehyde
Permeabilization Yes - eBioscience Permeabilization Buffer
Sample Human Cell (Differentiated Cardiac Ventricular Cells)
Specification Differentiated Cardiac Ventricular Cells
Gating Strategy Isotype negative control (white)
Preparation Cell harvesting/tissue preparation method: Typsin-EDTA Cell Dissociation
Sample buffer: PBS and 10% FBS
Username

Abcam user community

Verified customer

Submitted Feb 18 2014

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step Protein-Block Serum Free (DakoCytomation) as blocking agent as blocking agent for 10 minute(s) · Concentration: 0.25% · Temperature: RT°C
Antigen retrieval step None
Sample Sheep Tissue sections (Sheep tibia bone)
Specification Sheep tibia bone
Permeabilization No
Fixative 10% neutral-buffered formalin
Username

Abcam user community

Verified customer

Submitted Jun 18 2013

Thank you for your recent telephone enquiry.

As discussed on the telephone, far as we are aware, ab10288 Podoplanin / gp36 antibody [18H5] has never been tested with samples from mouse. All tested species cross-reactivity information is stat...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (thymus specimen)
Specification thymus specimen
Fixative Formaldehyde
Antigen retrieval step Heat mediated
Blocking step protein block as blocking agent for 5 minute(s) · Concentration: 0,015 mol/
Username

Dr. anca cimpean

Verified customer

Submitted Feb 02 2007

Thank you for your enquiry. The recommended method for this antibody is heat-mediated antigen retrieval with citrate buffer, pH 6.0. I have updated the datasheet with this information. I hope this helps. Please do not hesitate to contact us if ...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (dermal lymphatic endothelial)
Specification dermal lymphatic endothelial
Fixative Formaldehyde
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% goat
Username

Dr. Maria Navarro

Verified customer

Submitted May 03 2006

Thank you for your enquiry. I can confirm for you that this antibody, ab10288, is indeed a mouse monoclonal antibody (clone number 18H5). The statement in the relevance section, "Produced from sera of rabbits immunised with the recombinant ectodoma...

Read More

Thank you for your patience. We would like to confirm that this antibody has been tested on paraffin embedded section.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"