VE-Cadherin Antibody (BV14) - BSA Free

Western Blot: VE-Cadherin Antibody (BV14) [NBP1-43347] - Non-reduced (left) and reduced (right) bEnd.3 cell line lysates were loaded at 1x10^5 cells/lane, probed with 2 ug/mL of Anti-Mouse CD144 (VE-Cadherin) Purified and revealed with Anti-Rat IgG HRP.

MAP4K4 deficiency induces TGF beta /smad signaling and EndMT via activation of integrin beta 1. (a) HMVECs were transfected with MAP4K4 siRNA (100 nM) for 48 h. Next, the cells were treated with or without AcSDKP for 2 h. The p-smad3/smad3 pathway was analyzed by western blot. Densitometric analysis of the p-smad3/smad3 levels was performed, with n=3 for each group. (b) HMVECs were treated with MAP4K4 siRNA for 48 h with or without AcSDKP treatment. The VE-cadherin, CD31, FSP1, SM22 alpha and vimentin protein levels were analyzed by western blot. (c) HMVECs were transfected with MAP4K4 siRNA for 48 h in the presence or absence of TGF beta 2 with or without AcSDKP. The integrin beta 1 level was analyzed by western blot Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/28771231), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

AcSDKP suppresses TGF beta /smad signaling and EndMT through the FGFR1/FRS2 pathway. (a) HMVECs were treated with N-FGFR1 for 48 h, and the FGFR1, TGF beta R1 and TGF beta R2 protein levels were analyzed by western blot. (b) HMVECs were treated with TGF beta 2 in the presence or absence of N-FGFR1 for 15 min with or without AcSDKP preincubation. The p-smad3 and TGF beta R1 protein levels were analyzed by western blot. Densitometric analysis of the p-smad3/smad3 and TGF beta R1/ beta -actin levels (n=3) in each group was performed. (c) HMVECs were incubated with either N-FGFR1 in the presence or absence of TGF beta 2 for 48 h with or without preincubation with AcSDKP for 2 h or with N-FGFR1 in the presence or absence of TGF beta 2 for 48 h with or without 24 h of incubation with FGF2 (50 ng/ml). The CD31, SM22 alpha , FSP1 and alpha -SMA protein levels were analyzed by western blot. (d) HMVECs were transfected with FRS2 siRNA (100 nM) for 48 h with or without AcSDKP preincubation. The VE-cadherin, FSP1, vimentin, SM22 alpha and p-smad3 levels were analyzed by western blot. (e) HMVECs were treated with N-FGFR1 for 48 h or 15 min in the presence or absence of N-TGF beta (1, 2, 3) (1.0 μg/ml). The CD31, VE-cadherin, SM22 alpha , FSP1, TGF beta R1, TGF beta R2 and p-smad3 levels were analyzed by western blot Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/28771231), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

AcSDKP suppresses TGF beta /smad signaling and EndMT through the FGFR1/FRS2 pathway. (a) HMVECs were treated with N-FGFR1 for 48 h, and the FGFR1, TGF beta R1 and TGF beta R2 protein levels were analyzed by western blot. (b) HMVECs were treated with TGF beta 2 in the presence or absence of N-FGFR1 for 15 min with or without AcSDKP preincubation. The p-smad3 and TGF beta R1 protein levels were analyzed by western blot. Densitometric analysis of the p-smad3/smad3 and TGF beta R1/ beta -actin levels (n=3) in each group was performed. (c) HMVECs were incubated with either N-FGFR1 in the presence or absence of TGF beta 2 for 48 h with or without preincubation with AcSDKP for 2 h or with N-FGFR1 in the presence or absence of TGF beta 2 for 48 h with or without 24 h of incubation with FGF2 (50 ng/ml). The CD31, SM22 alpha , FSP1 and alpha -SMA protein levels were analyzed by western blot. (d) HMVECs were transfected with FRS2 siRNA (100 nM) for 48 h with or without AcSDKP preincubation. The VE-cadherin, FSP1, vimentin, SM22 alpha and p-smad3 levels were analyzed by western blot. (e) HMVECs were treated with N-FGFR1 for 48 h or 15 min in the presence or absence of N-TGF beta (1, 2, 3) (1.0 μg/ml). The CD31, VE-cadherin, SM22 alpha , FSP1, TGF beta R1, TGF beta R2 and p-smad3 levels were analyzed by western blot Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/28771231), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Spike-induced degradation of endothelial junctional proteins is greater in arteries of diabetic mice. (A) Representative Western blot indicating expression of endothelial junctional proteins in arteries isolated from control or diabetic mice. (B) Mean data indicating fold change in protein expression. n = 4 for each, *P < 0.05 vs. untreated control, δP < 0.05 vs. untreated diabetic, #P < 0.05 vs. Spike-treated control. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/34179146), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Either FGF19 or FGF21 enhanced the inhibitory effect of AcSDKP on EndMT and MEK/ERK pathway. In the presence of AcSDKP, HMVECs were stimulated by TGF beta 2 with or without FGF19 (100 ng·mL−1) or FGF21 (100 ng·mL−1) treatment. The levels of VE‐cadherin/ beta ‐Actin, alpha ‐SMA/ beta ‐Actin, P‐ERK/T‐ERK, and P‐MEK/T‐MEK were examined by western blot analysis (A, C) and quantified (B, D). The data represent mean +/- SD. Three independent experiments were performed for each result. ANOVA with Tukey's multiple comparisons test was applied. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/30972974), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

KLB deficiency led to EndMT in HMVECs. (A) Subconfluent HMVECs were transfected with KLB siRNA or control siRNA. Six hours later, the medium was replaced with an experimental medium, followed by N‐FGFR1 (1.5 mg·mL−1) treatment. At 48 h, the cells were harvested for western blot analysis. The results are from three repeated experiments. (B) The same treated HMVECs (as in A) cultured on 8‐well culture slides were subjected to immunofluorescence staining with an anti‐ alpha ‐SMA antibody and DAPI (scale bar, 100 μm). Six different fields were observed for each slide. (C) HMVECs with or without preincubation with AcSDKP (100 nm) for 2 h were transfected with KLB siRNA or control siRNA for 48 h. The expression of EndMT markers, including VE‐cadherin, alpha ‐SMA, vimentin, and SM22 alpha , was assessed by western blotting and quantified (D) by imagej software (GE Healthcare Life Sciences, Uppsala, Sweden). The data represent mean +/- SD and are representative of three independent experiments. One‐way ANOVA with Tukey's multiple comparisons test was used for statistical analysis. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/30972974), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

SARS-CoV-2 spike protein (Spike) increases endothelial permeability by downregulation of junctional proteins in diabetic endothelial cells. (A) Endothelial transwell permeability assay, mean data expressed as fold change. n = 6 for each, *P < 0.05 vs. untreated control, δP < 0.05 vs. untreated diabetic, #P < 0.05 vs. Spike-treated control. (B) Representative Western blot indicating expression of endothelial junctional proteins in endothelial cell culture with or without Spike treatment. (C) Mean data indicating fold change in protein expression. n = 6 for each, *P < 0.05 vs. untreated control, δP < 0.05 vs. untreated diabetic, #P < 0.05 vs. Spike-treated control. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/34179146), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Either FGF19 or FGF21 enhanced the inhibitory effect of AcSDKP on EndMT and MEK/ERK pathway. In the presence of AcSDKP, HMVECs were stimulated by TGF beta 2 with or without FGF19 (100 ng·mL−1) or FGF21 (100 ng·mL−1) treatment. The levels of VE‐cadherin/ beta ‐Actin, alpha ‐SMA/ beta ‐Actin, P‐ERK/T‐ERK, and P‐MEK/T‐MEK were examined by western blot analysis (A, C) and quantified (B, D). The data represent mean +/- SD. Three independent experiments were performed for each result. ANOVA with Tukey's multiple comparisons test was applied. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/30972974), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

• Reactivity: Hu, Mu
• Applications: WB, ICC/IF, IHC, IP
• Clone: BV14
• Clonality: Monoclonal
• Host: Rat
• Conjugate: Unconjugated
• Format: BSA Free
• Concentration: 0.5 mg/ml

VE-Cadherin Antibody (BV14) - BSA Free Summary

• Immunogen: This antibody is made against mouse VE-Cadherin
• Marker: Endothelial Cell Marker
• Isotype: IgG2b
• Clonality: Monoclonal
• Host: Rat
• Gene: CDH5
• Purity: Protein A or G purified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry/ Immunofluorescence
  • Immunohistochemistry 1:10-1:500
  • Immunohistochemistry-Frozen 1:10-1:500
  • Immunoprecipitation 1:10-1:500
  • Western Blot 2 ug/ml
• Application Notes: Use in WB reported in scientific literature (PMID:34179146)

Reactivity Notes

Use in Mouse reported in scientific literature (PMID:34179146).

Packaging, Storage & Formulations

• Storage: Store at 4C. Do not freeze.
• Buffer: PBS (pH 7.2)
• Preservative: 0.09% Sodium Azide
• Concentration: 0.5 mg/ml
• Purity: Protein A or G purified

Alternate Names for VE-Cadherin Antibody (BV14) - BSA Free

• 7B4 antigen
• 7B4
• cadherin 5, type 2 (vascular endothelium)
• cadherin 5, type 2, VE-cadherin (vascular epithelium)
• Cadherin-5
• CD144 antigen
• CD144
• CDH5
• endothelial-specific cadherin
• FLJ17376
• Vascular endothelial cadherin
• VECadherin
• VE-Cadherin

Background

The BV14 monoclonal antibody reacts with mouse VE-Cadherin (CD144). VE-Cadherin is a 120 kDa member of the type II Cadherin family, characterized by the presence of 5 extracellular cadherin domains (ECD), and anchored to the actin cytoskeleton through their cytoplasmic tail. VE-Cadherin mediates homophilic adhesion between neighbouring endothelial cells and is localized within specialized structures at cell-cell contacts, called adherens junctions. VE-Cadherin is expressed constitutively throughout the entire vasculature, and is required for numerous endothelial cell functions including migration, survival, contact-dependent growth inhibition and endothelial cell assembly into tubular structures. Furthermore, it is thought that VE-Cadherin+CD45- cells from the yolk sac or aorta-gonad-mesonephros (AGM)+ have the potential to give rise to hematopoietic cells. Cross-blocking experiments suggest that BV14 recognizes a different epitope than another mouse VE-Cadherin monoclonal antibody, BV13.

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 VE-Cadherin Antibody (NBP1-43347)(7)

Publications using NBP1-43347

• Raghavan S, Kenchappa DB,Leo MD SARS-CoV-2 Spike Protein Induces Degradation of Junctional Proteins That Maintain Endothelial Barrier Integrity Frontiers in cardiovascular medicine 2021-06-11 [PMID: 34179146] (WB, Mouse)
• Gao R, Kanasaki K, Li J et al. Beta klotho is essential for the anti-endothelial mesenchymal transition effects of N-acetyl-seryl-aspartyl-lysyl-proline FEBS Open Bio. [PMID: 30972974] (WB, ICC/IF, Human, Mouse)
• Li J, Shi S, et al. FGFR1 is critical for the anti-endothelial mesenchymal transition effect of N-acetyl-seryl-aspartyl-lysyl-proline via induction of the MAP4K4 pathway. Cell Death Dis 2017-08-03 [PMID: 28771231] (WB, Rabbit)
• Luo M, Flood EC, Almeida D et al. Annexin A2 supports pulmonary microvascular integrity by linking vascular endothelial cadherin and protein tyrosine phosphatases J. Exp. Med. 2017-07-10 [PMID: 28694388] (WB, Mouse)
• Crosby CV, Fleming PA, Argraves WS, Corada M, Zanetta L, Dejana E, Drake CJ. VE-cadherin is not required for the formation of nascent blood vessels but acts to prevent their disassembly. Blood;105(7):2771-6. 2005-04-01 [PMID: 15604224]
• Liao F, Li Y, O'Connor W, Zanetta L, Bassi R, Santiago A, Overholser J, Hooper A, Mignatti P, Dejana E, Hicklin DJ, Bohlen P. Monoclonal antibody to vascular endothelial-cadherin is a potent inhibitor of angiogenesis, tumor growth, and metastasis. Cancer Res;60(24):6805-10. 2000-12-15 [PMID: 11156369]
• Corada M, Mariotti M, Thurston G, Smith K, Kunkel R, Brockhaus M, Lampugnani MG, Martin-Padura I, Stoppacciaro A, Ruco L, McDonald DM, Ward PA, Dejana E. Vascular endothelial-cadherin is an important determinant of microvascular integrity in vivo. Proc Natl Acad Sci U S A;96(17):9815-20. 1999-08-17 [PMID: 10449777] (IF/IHC)

Bioinformatics

• Gene Symbol: CDH5
• Entrez:
  • 12562(Mouse)
• Uniprot:
  • P55284(Mouse)