Antibodies for Angiogenesis Research: LYVE-1, Prox-1, Podoplanin and CD31 (PECAM-1) - FocusOn 035

The circulation system consists of blood vessels and lymphatic vessels. The lymphatic system plays important roles in immune function, homeostasis and tumor metastasis. Clinical observations indicate thatwhile sarcomas spread mainly through the blood stream, most carcinomas spread primarily via the lymphatic vessels. In animal models, overexpression of lymphangiogenic factors VEGF-C and VEGF-D in tumor cells enhances metastatic spread of tumors. In the past, there were no reliable markers to distinguish blood vessel endothelial cells and lymphatic endothelial cells. Only recently several specific lymphatic markers have been discovered and „lymphangiogenesis“ is now emerging as an exciting sector of angiogenesis research, largely due to potential clinical applications. Acris Antibodies supports this research with a collection of well-validated antibodies against human and mouse lymphatic markers podoplanin, PROX-1 and LYVE-1.

Podoplanin

Podoplanin is a well-recognized lymphatic endothelium marker, which can be used to reliably distinguish lymphatic vessels from blood vessels. Podoplanin is a Mr 38000 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 occur 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.

Double Immunostaining of Human lymphatic endothelial cells using antibodies to Podoplanin (green, Cat.-No. DM3500) and PROX-1 (red)
Fig.1: Double Immunostaining of human lymphatic endothelial cells using antibodies to Podoplanin (green, Cat.-No. DM3500) and PROX-1 (red).
Fig. 2: Immunostaining of human mucosa (paraffin sections) using DM3500 (anti Podoplanin, left) and anti Prox-1 (right)
Fig.2: Immunostaining of human mucosa (paraffin sections) using anti Podoplanin Cat.-No. DM3500 (left) and anti Prox-1 (right)

LYVE-1

LYVE-1 has been identified as a major receptor for HA (extracellular matrix glycosamino-glycan hyaluronan) on the lymph vessel wall. The deduced amino acid sequence of LYVE-1 predicts a 322-residue type I integral membrane polypeptide 41% similar to the CD44 HA receptor with a 212-residue extracellular domain containing a single link module, the prototypic HA binding domain of the link protein superfamily. Like CD44, the LYVE-1 molecule binds both soluble and immobilized HA. However, unlike CD44, the LYVE-1 molecule colocalizes with HA on the luminal face of the lymph vessel wall and is absent from blood vessels. Hence, LYVE-1 is the first lymph-specific HA receptor to be characterized and is a uniquely powerful marker for lymph vessels themselves.

PROX-1

The homeobox gene PROX-1 was originally cloned by homology to the Drosophila melanogaster gene prospero. PROX-1 is expressed in a subpopulation of endothelial cells which by budding and sprouting, give rise to the lymphatic system. PROX-1-deficient homozygous mice have unaffected vasculogenesis and angiogenesis, but liver and lymphatics do not develop. Human PROX-1 gene has been mapped to chromosome 1q32.2- q32.3, and its product shows 94% similarity to the chicken protein. The conserved structure and expression pattern of the Prox-1 gene imply that it may play the same roles in humans as in other vertebrates. The human PROX-1 gene is most actively expressed in the developing lens. Acris Antibodies anti-PROX-1 provide a highly specific marker of lymphatic endothelial cells.

Antibody tools for studying lymphatic vessels

The combination of anti podoplanin, anti-LYVE-1 and anti-CD31 antibodies can be used to differentiate clearly between blood vessels, lymphatic vessels and bile ducts in paraffin embedded or frozen sections (see schema below). Furthermore they are useful for immunoprecipitation experiments and western blotting.

 PodoplaninCD31
Lymphatic vessel++
Blood vessel-+
Bile duct+-
Paraffin sections of mouse liver stained with Cat.-No. DM3501 (anti mouse Podoplanin, left) and anti mouse CD31 (right). Note: lymphatic vessels are Podoplanin+CD31+, blood vessels are Podoplanin- CD31+, bile ducts are Podoplanin+ CD31-.
Fig.3: Paraffin sections of mouse liver stained with Cat.-No. DM3501 (anti mouse Podoplanin, left) and anti mouse CD31 (right). Note: lymphatic vessels are Podoplanin+CD31+, blood vessels are Podoplanin- CD31+, bile ducts are Podoplanin+ CD31-.
Immunohistochemical staining on cryosections from human colon carcinoma using anti-human LYVE-1 antibody DP3500 at 10 µg/ml (pictures courtesy of Dr. U. Fiedler, S. Koidel, Tumor Biology Center, Freiburg, Germany).
Fig.4: Immunohistochemical staining on cryosections from human colon carcinoma using anti-human LYVE-1 antibody Cat.-No. DP3500 at 10 µg/ml (pictures courtesy of Dr. U. Fiedler, S. Koidel, Tumor Biology Center, Freiburg, Germany).
FACS analysis of LYVE-1 on human microvascular endothelial cells using antibody DP3500PS.
Fig.5
Immunohistochemical staining on paraffin sections of human intestine (border area of a colon carcinoma) using LYVE-1 antibody DP3500PS (picture courtesy of Dr. Karsten Debel, DCS, Hamburg, Germany).
Fig.6
Fig.5: FACS analysis of LYVE-1 on human microvascular endothelial cells using antibody Cat.-No. DP3500PS.
Fig.6: Immunohistochemical staining on paraffin sections of human intestine (border area of a colon carcinoma) using LYVE-1 antibody Cat.-No. DP3500PS (picture courtesy of Dr. Karsten Debel, DCS, Hamburg, Germany).

Product Citations (Selection)

Catalog-No.Citations
DM35001
DM35012
DP35003
DP35134
BM40475
BM40866
1
Kerstin Birke et al.: Expression of podoplanin and other lymphatic markers in the human anterior eye segment. Invest Ophthalmol Vis Sci, Jan 2010; 51(1): 344-354. PMID: 19737877
1
Joseph T. Turner et al.: Wolbachia lipoprotein stimulates innate and adaptive immunity through Toll-like receptors 2 and 6 to induce disease manifestations of filariasis. J Biol Chem, Aug 2009; 284(33): 22364-22378. PMID: 19458089
1
Nerbil Kilic et al.: Lymphatic reprogramming of microvascular endothelial cells by CEA-related cell adhesion molecule-1 via interaction with VEGFR-3 and Prox1. Blood, Dec 2007; 110: 4223-4233. PMID: 17761831
1
Rainer Riesenberg et al.: Expression of indoleamine 2,3-dioxygenase in tumor endothelial cells correlates with long-term survival of patients with renal cell carcinoma. Clin Cancer Res, Dec 2007; 13: 6993-7002. PMID: 18056175
1,4
Claudius Conrad et al.: Multipotent mesenchymal stem cells acquire a lymphendothelial phenotype and enhance lymphatic regeneration in vivo. Circulation, Jan 2009; 119: 281-289. PMID: 19118255
2
Pavel Uhrin et al.: Novel function for blood platelets and podoplanin in developmental separation of blood and lymphatic circulation. Blood, May 2010 (13); 115(19): 3997-4005. PMID: 20110424
2
Gabriela D’Amico et al.: Regulation of lymphatic blood vessel separation by endothelial Rac1. Development, Dec 2009; 136: 4043-4053. PMID: 19906871
2
Rolf Gräbner et al.: Lymphotoxin β receptor signaling promotes tertiary lymphoid organogenesis in the aorta adventitia of aged ApoE-/-mice. Exp Med, Jan 2009; 206: 233-248. PMID: 19139167
2
Masashi Gohda et al.: Sphingosine 1-phosphate regulates the egress of IgA plasmablasts from Peyer’s patches for intestinal IgA responses. J Immunol, Apr 2008; 180: 5335-5343. PMID: 18390715
3
Hiroshi Kimura et al.: Distinctive localization and opposed roles of vasohibin-1and vasohibin-2 in the regulation of angiogenesis. Blood, 2009 May 7;113(19):4810-4818. PMID: 19204325
3
Hans J. Geissler et al.: First year changes of myocardial lymphatic endothelial markers in heart transplant recipients. Eur J Cardiothorac Surg, May 2006; 29: 767-771. PMID: 16439147
4
Takahiro Heishi et al.: Endogenous angiogenesis inhibitor vasohibin1 exhibits broad-spectrum antilymphangiogenic activity and suppresses lymph node metastasis. Am J Pathol, Apr 2010; 176: 1950-1958. PMID: 20133819
5
Eric Soupene et al.: Characterization of an acyl-coenzyme A binding protein predominantly expressed in human primitive progenitor. J Lipid Res, May 2008; 49: 1103-1112. PMID: 18268358
6
Takayuki Morikawa et al.: Hypoxic regulation of the cerebral microcirculation is mediated by a carbon monoxide-sensitive hydrogen sulfide pathway.Proc Natl Acad Sci U S A, Jan 2012 (24); 109(4): 1293-1298. PMID: 22232681
6
Ji-Liang Li et al.: Delta-like 4 notch ligand regulates tumor angiogenesis, improves tumor vascular function, and promotes tumor growth in vivo. Cancer Res, Dec 2007; 67(23): 11244-11253. PMID: 18056450
6
Patrizia Larghero et al.: Biological assays and genomic analysis reveal lipoic acid cell behavior and gene expression modulation of endothelial cell behavior and gene expression. Carcinogenesis, May 2007; 28 (5): 1008–20. PMID: 17127716

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Primary Antibodies

Catalog No. Host Iso. Clone Pres. React. Applications  

CD31 / PECAM1 antibody

Immunohistochemical staining on Mouse Kidney Frozen Sections using CD31 antibody clone ER-MP12 Cat.-No BM4086B. Rat IgG2a ER-MP12 Purified Ms C, F, P
0.1 mg / €480.00
  Acris Antibodies GmbH

CD31 / PECAM1 (C-term) antibody

Human Cervix stained with anti-CD31 antibody Cat.-No AM11103PU Rabbit IgG SP38 Supernatant Hu, Ms P
0.1 ml / €290.00
  Acris Antibodies GmbH

CD31 / PECAM1 (C-term) antibody

AP15436PU CD31 antibody staining of Human Tonsil. Rabbit Aff - Purified Hu, Ms, Por P
0.1 ml / €180.00
  Acris Antibodies GmbH

CD31 / PECAM1 antibody

Figure 3. BM4047 CD31 antibody staining of Human Placenta Paraffin Section. Mouse IgG1 MEM-05 Hu C, F, P
0.2 mg / €310.00
  Acris Antibodies GmbH

CD31 / PECAM1 antibody

Staining of FFPE Human colon cancer (10x and 40x) with CD31/PECAM1 Antibody Cat.-No AM32708PU-N (Clone BC16-6.4EF) at 2 µg/ml. Antibody positive in cytoplasm of interstitial endotelial cells modulating neoangiogenesis. Mouse IgG1 BC16-6.4EF Purified Hu, Ms, Rt C, F, FN, ICC/IF, P, WB
0.1 mg / €250.00
  Acris Antibodies GmbH

CD31 / PECAM1 antibody

Western blot analysis of CD31 in THP-1 lysate probed with CD31 Antibody (Clone C31.7).  Mouse IgG1 C31.7 Purified Hu, Mky, Rb C, E, F, ICC/IF, IP, P, WB
0.1 mg / €320.00
  Acris Antibodies GmbH

CD31 / PECAM1 antibody

CD31 / PECAM1 Hamster IgG 2H8 Azide Free Ms F, FN
0.5 mg / €760.00
  Acris Antibodies GmbH

CD31 / PECAM1 antibody

CD31 / PECAM1 Mouse IgG1 TLD-3A12 Purified Rt C, E, F, ICC/IF, P, WB
0.1 mg / €260.00
  Acris Antibodies GmbH

LYVE-1 antibody

DP3500 LYVE-1 antibody staining of Human Spleen Paraffin section.
The experiment was performed by Prof. Dr. Birte Steiniger, Institute of Anatomy and Cell Biology Robert-Koch-Str. 8, D-35037 Marburg, Germany. Rabbit Purified Hu C, E, F, P, WB
0.1 mg / €270.00
  Acris Antibodies GmbH

LYVE-1 antibody

LYVE-1 antibody staining of Human Intestine (border area of a Colon Carcinoma).
The experiment has been performed by Dr. Karsten Debel, DCS, Hamburg, Germany. Rabbit Aff - Purified Hu C, E, F, P, WB
50 µg / €280.00
  Acris Antibodies GmbH

LYVE-1 antibody

Figure 4. LYVE-1 and CD31 staining on frozen sections (5μm) of the Mouse prostate.The experiments were performed by Scott Gerber & Edith Lord, PhD, University of Rochester, USA. Rabbit Purified Ms C, E, F, ICC/IF, WB
0.2 mg / €370.00
  Acris Antibodies GmbH

LYVE-1 antibody

Paraffin Section of Mouse Intestine stained with LYVE-1 Antibody Cat.-No DP3513P. You see the Staining (red) of lymphatic endothelial cells of the intestine. Rabbit IgG Aff - Purified Ms, Rt C, E, F, ICC/IF, P, WB
50 µg / €280.00
  Acris Antibodies GmbH

LYVE-1 antibody

DP3500 LYVE-1 antibody staining of Human Spleen Paraffin section.
The experiment was performed by Prof. Dr. Birte Steiniger, Institute of Anatomy and Cell Biology Robert-Koch-Str. 8, D-35037 Marburg, Germany. Rabbit Purified Hu C, E, F, P, WB
0.2 mg / €370.00
  Acris Antibodies GmbH

LYVE-1 antibody

Figure 4. LYVE-1 and CD31 staining on frozen sections (5μm) of the Mouse prostate.The experiments were performed by Scott Gerber & Edith Lord, PhD, University of Rochester, USA. Rabbit Purified Ms C, E, F, ICC/IF, WB

Regular Price: 0.1 mg / €270.00

Special Price: 0.1 mg / €202.50

  Acris Antibodies GmbH

LYVE-1 (25-235) antibody

Western blot analysis: A549 (lung carcinoma) cell lysate was resolved by SDS-PAGE, transferred to PVDF membrane and probed with anti-human LYVE-1 antibody (1/500). Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP and an ECL detection system. Mouse IgG2b 4G1 Purified Hu E, F, WB
50 µl / €230.00
  Acris Antibodies GmbH

Podoplanin antibody

DM3500 Podoplanin antibody staining of Paraffin Embedded Human Mucosa Sections. Mouse IgG1 18H5 Supernatant Hu C, F, IP, P, WB
0.2 ml / €370.00
  Acris Antibodies GmbH

Podoplanin antibody

Immunofluorescence staining (green) of Podoplanin in primary human dermal lymphatic endothelial cells (HDLEC) with anti-Human Podoplanin Antibody VCat.-No DM3500P (0.5 μg/ml) and counter staining of nuclei with Dapi. As secondary antibody Goat anti-Mouse ALEXA Fluor 488 was used 1/400. Mouse IgG1 18H5 Aff - Purified Hu C, F, ICC/IF, P, WB
0.1 mg / €350.00
  Acris Antibodies GmbH

Podoplanin antibody

NZ-1 Staining of Human Mesothelial Cells. Rat IgG2a NZ-1 Purified Hu C, F, IP, P, WB
100 µg / €370.00
  Acris Antibodies GmbH

Podoplanin antibody

Immunohistochemistry of Mouse Liver (paraffin embedded sections) using anti-Podoplanin antibody Hamster IgG 811 Purified Ms C, IP, P, WB
0.1 mg / €340.00
  Acris Antibodies GmbH

Podoplanin antibody

Western blot analysis using Podoplanin Antibody Cat.-No DM3516P with recombinant Human, Mouse and Rat soluble Podoplanin. There is no cross reaction with Human Podoplanin. Hamster IgG RTD4E10 Purified Ms C, IP, P, WB
0.1 mg / €350.00
  Acris Antibodies GmbH

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