Anti-Podoplanin antibody [18H5] - BSA and Azide free (ab10288)
Key features and details
- Mouse monoclonal [18H5] to Podoplanin - BSA and Azide free
- Suitable for: ICC/IF, WB, Flow Cyt
- Reacts with: Human
- Isotype: IgG1
Related conjugates and formulations
Overview
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Product name
Anti-Podoplanin antibody [18H5] - BSA and Azide free
See all Podoplanin primary antibodies -
Description
Mouse monoclonal [18H5] to Podoplanin - BSA and Azide free -
Host species
Mouse -
Tested applications
Suitable for: ICC/IF, WB, Flow Cytmore details -
Species reactivity
Reacts with: Human -
Immunogen
Full length protein corresponding to Podoplanin. MDCK cells stably expressing Podoplanin protein.
Database link: Q86YL7 -
Positive control
- WB: human lymphatic endothelial cells, human placenta and skeletal muscle tissue lysates. ICC/IF: human dermal lymphatic endothelial Flow Cyt: HCT 116 (Human colorectal carcinoma cell line) whole cell lysate,
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General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Properties
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Form
Liquid -
Storage instructions
Shipped 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 buffer
Constituent: PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Protein G purified -
Clonality
Monoclonal -
Clone number
18H5 -
Isotype
IgG1 -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
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 |
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ICC/IF | (1) |
Use a concentration of 1 - 2 µg/ml.
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WB | (1) |
Use a concentration of 5 µg/ml. Detects a band of approximately 35 kDa (predicted molecular weight: 24 kDa).
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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.
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Notes |
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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.
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Target
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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
modificationsExtensively 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
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Database links
- Entrez Gene: 10630 Human
- Omim: 608863 Human
- SwissProt: Q86YL7 Human
- Unigene: 468675 Human
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Alternative names
- Aggrus antibody
- D2-40 antibody
- Glycoprotein 36 KD antibody
see all
Images
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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.
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All lanes : Anti-Podoplanin antibody [18H5] - BSA and Azide free (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.
Secondary
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. -
Flow cytometric analysis of human primary lymphatic endothelial cells (HDLEC) labeling Podoplanin with ab10288 at 5 µg/mL. Anti-mouse IgG-PE was used as a secondary antibody.
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Flow cytometric analysis of primary human dermal microvascular (HDMVEC) and umbilical vein (HUVEC) endothelial cells labeling Podoplanin with ab10288 at 5 µg/mL. The lymphatic endothelial cell marker Podoplanin is not expressed in the blood endothelial cells.
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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.
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Immunocytochemistry/Immunofluorescence analysis of human dermal lymphatic endothelial cells (HDLEC) labelling Podoplanin with ab10288. An Alexa Fluor® 488-conjugated goat anti-mouse was used as the secondary antibody. Nuclei counterstained with DAPI.
Protocols
Datasheets and documents
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Datasheet download
References (41)
ab10288 has been referenced in 41 publications.
- Luks VL et al. Surface conjugation of antibodies improves nanoparticle uptake in bronchial epithelial cells. PLoS One 17:e0266218 (2022). PubMed: 35385514
- Wang S et al. Inflammatory macrophages interrupt osteocyte maturation and mineralization via regulating the Notch signaling pathway. Mol Med 28:102 (2022). PubMed: 36058911
- Li S et al. An integrated map of fibroblastic populations in human colon mucosa and cancer tissues. Commun Biol 5:1326 (2022). PubMed: 36463319
- Rosa I et al. The Soluble Guanylate Cyclase Stimulator BAY 41-2272 Attenuates Transforming Growth Factor β1-Induced Myofibroblast Differentiation of Human Corneal Keratocytes. Int J Mol Sci 23:N/A (2022). PubMed: 36499651
- Williams R et al. Age-Related Changes in Female Scalp Dermal Sheath and Dermal Fibroblasts: How the Hair Follicle Environment Impacts Hair Aging. J Invest Dermatol 141:1041-1051 (2021). PubMed: 33326808