Pdpn Hamster Monoclonal Antibody [Clone ID: 811]
Pdpn hamster monoclonal antibody, clone 811, Purified
|Applications||IHC, IP, WB|
|Recommended Dilution||Immunoprecipitation (1/50)
Immunohistochemistry on Frozen and Paraffin Embedded Sections (1/50-1/200).
Western blotting (≥ 1/100). The antibody will detect a 36-40 kDa band.
|Specificity||This Syrian Hamster monoclonal antibody will detect the extracellular domain of Podoplanin on the surface of lymphatic endothelial cells and some epithelial cell types by immunostaining or immunohistochemistry.
Negative Species: Human.
State: Liquid purified IgG fraction
|Reconstitution Method||The antibody can be reconstituted in 0.2 ml water.|
|Purification||Affinity Chromatography on Protein G|
|Storage||Store the antibody undiluted at 2-8°C for one month or (in aliquots) at -20°C for longer (add 50% Glycerol).
Avoid repeated freezing and thawing.
|Stability||Shelf life: one year from despatch.|
|Background||Podoplanin is a well-recognized lymphatic endothelium marker, which can be used to reliably distinguish lymphatic vessels from blood vessels. Podoplanin is a Mr 38,000 membrane mucoprotein that was originally detected on the surface of rat glomerular epithelial cells (podocytes) and was found to be linked to flattening of foot processes that occurs in glomerular diseases. Podoplanin shows features of a membrane mucoprotein with several conserved O-glycosylation sites. Currently, it is of unknown biological function. Because heavily O-glycosylated mucoproteins were identified recently as counterreceptors for selectins that mediate adhesion of inflammatory cells, it is possible that podoplanin plays a similar role in lymphatic endothelia.|
|Synonyms||Glycoprotein 36, PA2.26 antigen, T1-alpha, Aggrus, PDPN, GP36, PSEC0003, PSEC0025|
|Note||Protocol: IMMUNOSTAINING ON FORMALIN FIXED PARAFFIN-EMBEDDED SECTIONS
1. Deparaffinize through Xylenes, Ethanol , and hydrate to water.
2. 1% Hydrogen peroxide in Methanol , 30-40 min, RT.
3. Wash 2x, dd water, wash in Tris buffer, pH 7.2, 5 min. each.
4. Proteinase K in 0.2M Tris, buffer, pH 7.2, 37°C, 36 µg/ml, 30 min.
5. Wash 3x, 5 min each: Tris buffer, pH 7.2, dd sterile water; TNT wash buffer.
6. Block in TNB blocking buffer, 30 min, RT.
7. Primary antibody: Hamster anti-Mouse Podoplanin antibody 5-10 µg/ml in TNB buffer, overnight, 4°C.
8. Rinse in TNT wash buffer, 3x, 5 min.
9. Incubate with Rabbit anti-Syrian Hamster IgG HRP conjugated, 1:100 in TNB buffer, 30 min, RT. Wash 3x.
10. Biotinylated-tyramide in amplification diluent, 1:50, 7 min, RT Wash 3x.
11. Incubate in SA-HRP , 1:100 in TNB, 30 min, RT. Wash 3x.
13. Counter stain.