Recombinant Anti-Podoplanin antibody [EPR22182] (ab236529)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR22182] to Podoplanin
- Suitable for: WB, IHC-P, IP, Flow Cyt (Intra), Flow Cyt, mIHC
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-Podoplanin antibody [EPR22182]
See all Podoplanin primary antibodies -
Description
Rabbit monoclonal [EPR22182] to Podoplanin -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-P, IP, Flow Cyt (Intra), Flow Cyt, mIHCmore details -
Species reactivity
Reacts with: Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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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 (intra): U-87 MG and U-2 OS cells. IP: U-2 OS whole cell lysate. mIHC-P: Human lung tissue. Flow cyto: U-87 MG cells
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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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
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 long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR22182 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
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 |
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WB |
1/1000. Detects a band of approximately 38 kDa (predicted molecular weight: 16 kDa).
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IHC-P |
1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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IP |
1/30.
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Flow Cyt (Intra) |
1/300.
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Flow Cyt |
Use at an assay dependent concentration.
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mIHC |
1/8000.
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Notes |
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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. |
IP
1/30. |
Flow Cyt (Intra)
1/300. |
Flow Cyt
Use at an assay dependent concentration. |
mIHC
1/8000. |
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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Flow cytometry overlay histogram showing left U-87 MG positive cells and right negative HUVEC stained with ab236529 (red line). The cells were incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab236529) (1x 106 in 100μl at 1.0 μg/ml (1/500)) for 30min on ice.
The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min on ice
Isotype control antibody (black line) was Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.
Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.
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Fluorescence multiplex immunohistochemical analysis of formalin/PFA-fixed paraffin-embedded Human lung tissue.
Panel A: Merged staining of anti-Podoplanin (magenta; Opal™690), anti-Surfactant protein D/SP-D (green; Opal™520) and anti-Sialoadhesin/CD169 (red; Opal™570) on human lung.Panel B: Anti-Podoplanin stained on alveolar type I cells.
Panel C: Anti-Surfactant protein D/SP-D stained on alveolar type II cells.
Panel D: Anti-Sialoadhesin/CD169 stained on macrophages.The section was incubated in three rounds of staining: in the order of ab236529 and ab234257 for 30 mins, then ab245735 for 10 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins. Counterstained with DAPI.
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Immunohistochemical analysis of paraffin-embedded human colon tissue labeling Podoplanin 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.
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Podoplanin 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.
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Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized U-2 OS (human bone osteosarcoma epithelial cell line) cell line labeling Podoplanin 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.
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Immunohistochemical analysis of paraffin-embedded human mesothelioma tissue labeling Podoplanin 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.
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All lanes : Anti-Podoplanin 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 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
The molecular mass observed is consistent with what has been described in the literature (PMID: 10408868; PMID: 17279584).
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Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized U-87 MG (humanglioblastoma-astrocytoma epithelial cell line) cell line labeling Podoplanin 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.
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All lanes : Anti-Podoplanin 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 minutesBlocking/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).
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Immunohistochemical analysis of paraffin-embedded human Kaposi’s sarcoma tissue labeling Podoplanin 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.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (6)
ab236529 has been referenced in 6 publications.
- Wang X et al. PDPN contributes to constructing immunosuppressive microenvironment in IDH wildtype glioma. Cancer Gene Ther 30:345-357 (2023). PubMed: 36434176
- Lv X et al. The cell cycle inhibitor P21 promotes the development of pulmonary fibrosis by suppressing lung alveolar regeneration. Acta Pharm Sin B 12:735-746 (2022). PubMed: 35256943
- Rodriguez AB et al. Immune mechanisms orchestrate tertiary lymphoid structures in tumors via cancer-associated fibroblasts. Cell Rep 36:109422 (2021). PubMed: 34289373
- Wang S et al. A single-cell transcriptomic landscape of the lungs of patients with COVID-19. Nat Cell Biol 23:1314-1328 (2021). PubMed: 34876692
- Lin S et al. Prognosis Analysis and Validation of m6A Signature and Tumor Immune Microenvironment in Glioma. Front Oncol 10:541401 (2020). PubMed: 33123464
- Xiong HG et al. Long noncoding RNA MYOSLID promotes invasion and metastasis by modulating the partial epithelial-mesenchymal transition program in head and neck squamous cell carcinoma. J Exp Clin Cancer Res 38:278 (2019). PubMed: 31238980