Recombinant Anti-Podoplanin / gp36 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 / gp36
- Suitable for: WB, IHC-P, IP, Flow Cyt (Intra)
- Reacts with: Human
Related conjugates and formulations
Overview
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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, IP, Flow Cyt (Intra)more 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.
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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) |
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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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. |
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
- Glycoprotein 36 KD antibody
- Glycoprotein 36 antibody
see all
Images
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Podoplanin / gp36 antibody [EPR22182] (ab236529)
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.
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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.
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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 / 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.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Podoplanin / gp36 antibody [EPR22182] (ab236529)
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.
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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 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 / 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.
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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 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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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Podoplanin / gp36 antibody [EPR22182] (ab236529)
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.
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 (2)
ab236529 has been referenced in 2 publications.
- 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