Mouse Anti-Human Podoplanin
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Cat-Nr. | 101-M40 |
Size | 100 µg |
Price | 275 € |
Category | Monoclonal Antibody |
Clone Nr. | (#18H5) |
Isotype | IgG1 |
Species Reactivity | Human |
Formulation | lyophilized |
Buffer | PBS |
Reconstitution | Centrifuge vial prior to opening. Reconstitute in sterile water to a concentration of 0.1-1.0 mg/ml. |
Stability and Storage | The lyophilized antibody is stable for at least 2 years at -20°C. After sterile reconstitution the antibody is stable at 2-8°C for up to 6 months. Frozen aliquots are stable for at least 6 months when stored at -20°C. Addition of a carrier protein or 50% glycerol is recommended for frozen aliquots. |
Preparation | The monoclonal antibody was produced with the help of BALB/c mice using MDCK cells stably expressing gp36 (Podoplanin) protein as the immunizing antigen. The mouse IgG1 antibody (#18H5) from hybridomas was purified from cell culture supernatant by Protein G chromatography. |
Antigen | gp36 (Podoplanin)-expressing MDCK cells |
Application | WB, IHC, IF, FC |
Synonyms | PDPN; T1A; GP36; GP40; Gp38; OTS8; T1A-2; AGGRUS; HT1A-1; PA2.26 |
Description | Podoplanin, also known as glycoprotein 36 (gp36), PA2.26 antigen, T1alpha (T1A), and aggrus, is a 36 kDa type I transmembrane sialoglycoprotein and member of the Podoplanin family. Podoplanin has three potential splice variants, the longest of which is represented by a 238 amino acid (aa) precursor (NP_006465). It contains an undefined signal sequence, a 22 aa transmembrane segment (aa 207-228) and a short cytoplasmic tail (aa 229-238). The ECD contains abundant Ser/Thr residues that could serve as potential O-linked glycosolation sites. The cytoplasmic tail contains putative sites for protein kinase C phosphorylation. There are two potential alternate start sites at Met 77 (Swiss Prot #: Q86YL7) and Met 119 (EAW51692) that generate short forms. The 162 aa short form Podoplanin precursor shares 47% aa identity with mouse Podoplanin. Podoplanin is expressed on glomerular epithelial cells (podocytes), type I lung alveolar cells, lymphatic endothelial cells, and numerous tumors, including colorectal tumors, squamous cell carcinomas, testicular seminoma, and brain tumors. One study shows high expression of Podoplanin mRNA in placenta, lung, skeletal muscle, and heart, and weaker levels in brain, kidney, and liver. Podoplanin is the ligand for C-type lectin-like receptor 2 (CLEC2). Their association is dependent on sialic acid on O-glycans of Podoplanin. Through its association with CLEC2, Podoplanin-induces platelet aggregation and tumor metastasis. Podoplanin is also necessary for lymphatic vessel formation, normal lung cell proliferation and alveolus formation at birth. |
Uniprot ID | Q86YL7 |
Protein RefSeq | NP_001006625.1 |
mRNA RefSeq | NM_001006624.1 |
Figures
![](/fileadmin/user_upload/products/Podoplanin/101-M40_IF.jpg)
Immunofluorescence staining (green) of Podoplanin in primary human dermal lymphatic endothelial cells (HDLEC) with anti-human Podoplanin (0,5µg/ml) [Cat# 101-M40] and counter staining of nuclei with Dapi. As secondary antibody goat anti-mouse ALEXA Flour 488 (Dianova) was used 1:400.
![](/fileadmin/user_upload/products/Podoplanin/101-M40_IHC_1.jpg)
Immunohistochemistry on cryo sections of human spleen using a mouse anti-human Podoplanin antibody (Cat# 101-M40). The antibody stains solely the lymphatic endothelial cells and not the sinus endothelium.
Detection system: Vector Labs' Vectastain Elite system, used as the manufacturer provides.
Experiment was performed by Prof. Dr. Birte Steiniger, Institute of Anatomy and Cell Biology Robert-Koch-Str. 8, D-35037 Marburg, Germany
![](/fileadmin/user_upload/products/Podoplanin/101-M40_FACS_PE.jpg)
FACS analysis with human primary lymphatic endothelial cells (HDLEC). As secondary antibody anti-mouse IgG-PE was used.
![](/fileadmin/user_upload/products/Podoplanin/101-M40_WB_Lysate_red.png)
Western Blot Analysis with total lysate of different LEC cell lines using a monoclonal mouse anti-human Podoplanin antibody [Cat# 101-M40]. The proteins have been denaturated at 95°C for 5 min. The LEC cell lines HDLEC-c5 and LyLec12 showed no signal for Podoplanin expression.
![](/fileadmin/user_upload/products/Podoplanin/101-M40_FACS_HDMVEC_HUVEC.jpg)
FACS analysis with primary human dermal microvascular (HDMVEC) and umbilical vein (HUVEC) endothelial cells. The lymphatic endothelial cell marker Podoplanin is not expressed in the blood endothelial cells.
![](/fileadmin/user_upload/products/Podoplanin/101-M40_WB_Lysate_HUVEC_LEC.jpg)
Western blot analysis of Podoplanin expression in human lymphatic endothelial cells (LEC) and HUVECs. Total lysate of both cell types were subjected to SDS-PAGE and subsequent Western analysis with the monoclonal
![](/fileadmin/user_upload/products/Podoplanin/101-M40_WB_Lysate_non-red.png)
Western Blot Analysis with total lysate of different LEC cell lines and the neuroblastoma cell line Sy5y using a monoclonal mouse anti-human Podoplanin antibody [Cat# 101-M40]. The LEC cell line LyLec12 and the neuroblastoma cell line Sy5y showed no signal for Podoplanin expression. The SDS-PAGE was performed under non-reducing conditions.
Reference
- Heterogeneity of stromal cells in the human splenic white pulp. Fibroblastic reticulum cells, follicular dendritic cells and a third superficial stromal cell type. Steiniger B. et al., Immunology. 2014 Nov; 143(3): 462–477.
- Lack of Evidence for the Direct Activation of Endothelial Cells by Adult Female and Microfilarial Excretory-Secretory Products. T. Weinkopff and P. Lammie, PLoS One. 2011; 6(8): e22282.
- Aberrant lymphatic development in euploid fetuses with increased nuchal translucency including Noonan syndrome. de Mooij YM et al., Prenat Diagn. 2011 Feb;31(2):159-66.
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