Product nameAnti-Podoplanin / gp36 antibody [18H5]
See all Podoplanin / gp36 primary antibodies
DescriptionMouse monoclonal [18H5] to Podoplanin / gp36
Tested applicationsSuitable for: IHC-P, ICC/IF, WB, Flow Cyt, IHC-Frmore details
Species reactivityReacts with: Human
Full length protein corresponding to Podoplanin/ gp36. MDCK cells stably expressing Podoplanin/ gp36 protein.
Database link: Q86YL7
- In Western blot, this antibody gave a positive signal in human placenta and skeletal muscle tissue lysates.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Storage bufferpH: 7.40
Concentration information loading...
PurityProtein G purified
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.
|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)
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
|IHC-Fr||Use at an assay dependent concentration. PubMed: 18987360|
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.
modificationsExtensively 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
- Aggrus antibody
- Glycoprotein 36 KD antibody
- Glycoprotein 36 antibody
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.
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.
All lanes : Anti-Podoplanin / gp36 antibody [18H5] (ab10288) at 5 µg/ml
Lane 1 : Human placenta tissue lysate - total protein (ab29745)
Lane 2 : Human skeletal muscle tissue lysate - total protein (ab29330)
Lysates/proteins at 20 µg per lane.
All lanes : 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.
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.
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.
This product has been referenced in:
- Zhao X et al. Plasma soluble podoplanin is a novel marker for the diagnosis of tumor occurrence and metastasis. Cancer Sci 109:403-411 (2018). Read more (PubMed: 29266546) »
- Nikitakis NG et al. DSPP-MMP20 gene silencing downregulates cancer stem cell markers in human oral cancer cells. Cell Mol Biol Lett 23:30 (2018). Read more (PubMed: 30002682) »