Podocalyxin Like Antibody (3D3)

Western Blot: Podocalyxin Like Antibody (3D3) [NBP2-25219] - Western Blot Analysis of HeLa cell lysate using Podocalyxin Like Antibody (3D3).

Immunocytochemistry/Immunofluorescence: Podocalyxin Like Antibody (3D3) [NBP2-25219] - Confocal Immunofluorescence of HeLa cells using Podocalyxin Like Antibody (3D3) labeled with CF488 (Green); Red Dot is used to label the nuclei.

Immunohistochemistry-Paraffin: Podocalyxin Like Antibody (3D3) [NBP2-25219] - Formalin-fixed, paraffin-embedded human Placenta stained with Podocalyxin Like Antibody (3D3).

Flow Cytometry: Podocalyxin Like Antibody (3D3) [NBP2-25219] - Surface staining of NCCIT cells using Podocalyxin antibody at 1 ug/10^6 cells. A FITC-conjugated anti mouse Ig secondary antibody was used along with this antibody .

Immunohistochemistry-Paraffin: Podocalyxin Like Antibody (3D3) [NBP2-25219] - Formalin-paraffin angiosarcoma stained with Podocalyxin Ab (3D3).

Immunohistochemistry-Paraffin: Podocalyxin Like Antibody (3D3) [NBP2-25219] - Formalin-fixed, paraffin-embedded human Angiosarcoma stained with Podocalyxin Like Antibody (3D3).

• Reactivity: Hu, Mu, Rt, Rb
• Applications: WB, Flow, ICC/IF, IHC, IP
• Clone: 3D3
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Concentration: 0.2 mg/ml

Podocalyxin Like Antibody (3D3) Summary

• Description: 200ug/ml of antibody purified from Bioreactor Concentrate by Protein A or G. Prepared in 10 mM PBS with 0.05% BSA & 0.05% azide. Also available WITHOUT BSA & azide at 1.0 mg/ml. (NBP2-33108)
  
  Antibody with azide - store at 2 to 8C. Antibody without azide - store at -20 to -80C.
• Immunogen: A recombinant protein fragment containing the intracellular, transmembrane, and part of the extracellular domain of human Podocalyxin Like (Uniprot: O00592)
• Localization: Cell surface and Cytoplasmic
• Marker: Hematopoietic Stem Cell Marker
• Specificity: Podocalyxin is a member of the CD34 transmembrane sialomucin family. It is over-expressed on the podocyte foot projections and plays essential roles in kidney development and homeostasis, blood filtration and urine formation. It is also expressed on vascular endothelia, hematopoietic progenitors and a subset of neurons. Overexpression of podocalyxin may be linked to more aggressive tumor behavior. Podocalyxin antibody can identify podocytes in the urine (podocyturia) that may indicate glomerular disease, pre-eclampsia, and other kidney pathology.
• Isotype: IgG1 Kappa
• Clonality: Monoclonal
• Host: Mouse
• Gene: PODXL
• Purity: Protein A or G purified

Applications/Dilutions

• Dilutions:
  • Flow (Intracellular)
  • Flow Cytometry 1-2 ug/million cells
  • Immunocytochemistry/ Immunofluorescence 1-2 ug/ml
  • Immunohistochemistry
  • Immunohistochemistry-Frozen 0.5-1ug/ml
  • Immunohistochemistry-Paraffin 1-2 ug/ml
  • Immunoprecipitation 1-2 ug/500 ug protein
  • Western Blot 1-2 ug/ml
• Application Notes: Immunohistochemistry (Formalin-fixed): 1-2ug/ml for 30 min at RT. Staining of formalin-fixed tissues requires heating tissue sections in 10mM Tris with 1mM EDTA, pH 9.0, for 45 min at 95C followed by cooling at RT for 20 minutes.
  Optimal dilution for a specific application should be determined.

Reactivity Notes

Mouse reactivity reported in scientific literature (PMID: 27771514).

Packaging, Storage & Formulations

• Storage: Store at 4C.
• Buffer: 10 mM PBS with 0.05% BSA
• Preservative: 0.05% Sodium Azide
• Concentration: 0.2 mg/ml
• Purity: Protein A or G purified

Alternate Names for Podocalyxin Like Antibody (3D3)

• GCTM
• Gp200MGC138240
• PCLP
• PCLP1
• PCLP-1
• PCLPGCTM-2 antigen
• PCpodocalyxin
• POD XL
• Podocalyxin
• Podocalyxin-like protein 1
• podocalyxin-like
• PODXL

Background

Podocalyxin, also known as podocalyxin-like or PDXL, is a member of the CD34 transmembrane sialomucin family. Like other sialoglycoproteins, podocalyxin plays important roles in mediating cellular migration and cell-cell adhesion. Podocalyxin was first discovered in podocytes, the terminally differentiated epithelial cells lining the kidney glomeruli. Podocalyxin is expressed at high levels on the podocyte foot projections and plays essential roles in kidney development and homeostasis, particularly blood filtration and urine formation.
Podocalyxin is also expressed on vascular endothelia, hematopoietic progenitors and a subset of neurons (reviewed in Nielsen, 2009). Aberrant expression of podocalyxin has been described in a variety of cancers including breast, prostate, liver, pancreatic, kidney and leukemia. Human podocalyxin migrates as a 165/170 kDa doublet on SDS-PAGE; however there are many post-translational modifications which may impact its actual size (Kershaw, 1997).
Antibody to podocalyxin has been reported to be a useful biomarker for various pathological conditions. For example, podocalyxin antibody can identify podocytes in the urine, also known as podocyturia, which can be indicative of glomerular disease, pre-eclampsia, and other kidney pathology (reviewed in Chen, 2012). Additionally, podocalyxin antibody studies have indicated that highly podocalyxin-positive tumors or podocalyxin dysregulation in general can be potentially linked to more aggressive tumors and poorer outcomes (Nielson, 2009). Hence, podocalyxin antibody may be used as part of a biomarker antibody panel to study expression of proteins associated with tumorigenesis. The podocalyxin antibody clones 3D3, 4F10, and 2A4 have been extensively published in the literature and researchers are encouraged to consult the published literature for additional information.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Podocalyxin Like Antibody (NBP2-25219)(9)

Publications using NBP2-25219

• Dwivedi O, Barreiro K, Käräjämäki A et al. Genome-wide mRNA profiling in urinary extracellular vesicles reveals stress gene signature for diabetic kidney disease iScience 2023-04-01 [PMID: 37216114]
• Marthinus W Myburgh, Lorenzo Favaro, Willem H van Zyl, Marinda Viljoen-Bloom Engineered yeast for the efficient hydrolysis of polylactic acid. Bioresource technology 2023-04-18 [PMID: 37011843]
• Barreiro K, Dwivedi OP, Leparc G, et al. Comparison of urinary extracellular vesicle isolation methods for transcriptomic biomarker research in diabetic kidney disease Journal of extracellular vesicles 2020-12-01 [PMID: 33437407] (WB, Human)
• Yuvashree, M, Ganesh, R N Et al. Potential application of nanoemulsified garlic oil blend in mitigating the progression of type 2 diabetes-mediated nephropathy in Wistar rats. 3 Biotech 2020-06-01 [PMID: 32523866] (ELISA, Rat)
• Puhka M, Nordberg ME, Valkonen S et al. KeepEX, a simple dilution protocol for improving extracellular vesicle yields from urine. Eur J Pharm Sci 2016-10-19 [PMID: 27771514] (WB, Mouse)
• Siddiqi FS, Chen L, Advani SL et al. CXCR4 promotes renal tubular cell survival in male diabetic rats: implications for ligand inactivation in the human kidney. Endocrinology. 2014-12-30 [PMID: 25549045] (IHC-P, Human)
  
  Details:
  Podocalyxin antibody used for Immunohistochemistry on formalin-fixed paraffin-embdded human kidney tissue.
• Schopperle WM, Kershaw DB, DeWolf WC. Human embryonal carcinoma tumor antigen, Gp200/GCTM-2, is podocalyxin. Biochem Biophys Res Commun. 2003-01-10 [PMID: 12504081]
• Sassetti C, Tangemann K, Singer MS et al. Identification of podocalyxin-like protein as a high endothelial venule ligand for L-selectin: parallels to CD34. J Exp Med. 1998-06-15 [PMID: 9625756] (IHC-Fr, WB)
  
  Details:
  IHC (frozen): Fig 1 (tonsil), Fig 2 (tonsil, appendix, lymph node) WB: Fig 4A & 6 (purified podocalyxin from tonsil) IP: Fig 4B (purified posoxcalyxin from tonsil) Parallel plate flow chamber analysis: Fig 7 (lymphocytes, Jurkat). Refer to the publication
• Kershaw DB, Beck SG, Wharram BL et al. Molecular cloning and characterization of human podocalyxin-like protein. Orthologous relationship to rabbit PCLP1 and rat podocalyxin. J Biol Chem. 1997-06-20 [PMID: 9188463] (IHC-Fr, WB, IP, Human, Rat, Rabbit)
  
  Details:
  Podocalyxin Antibody clones cited: 2A4 (ABX-166A), 4F10 (ABX-165A), and 3D3 (ABX-164A): IHC (frozen): Fig 4 (kidney cortex) WB: Fig 5 (podocalyxin human and rabbit fusion proteins), Fig 7A-D (human, rabbit and rat renal glomerular extracts) IP: Fig 7E-F (

Bioinformatics

• Gene Symbol: PODXL
• Uniprot:
  • NP_001018121(Human)
  • O00592(Human)