Anti nrp 1

Manufactured by Santa Cruz Biotechnology

Anti-NRP-1 is a laboratory product that targets the Neuropilin-1 (NRP-1) protein. NRP-1 is a cell surface receptor involved in various cellular processes. This product can be used to detect and study the NRP-1 protein in research applications.

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Lab products found in correlation

Fv1000mpe confocal microscope, by Olympus (2 mentions) Anti cd31, by Santa Cruz Biotechnology (2 mentions) Anti cd144, by Thermo Fisher Scientific (2 mentions) Fv10 asw 3.0 viewer software, by Olympus (1 mentions) 4 6 diamidino 2 phenylindole dapi, by Agilent Technologies (1 mentions) Uplansapo 60xw, by Olympus (1 mentions) Anti β actin, by Cell Signaling Technology (1 mentions) Alexa 488, by Thermo Fisher Scientific (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions) Nitrocellulose membrane, by GE Healthcare (1 mentions) Emt antibody sampler kit, by Cell Signaling Technology (1 mentions) Anti pdgfrα, by Santa Cruz Biotechnology (1 mentions) Anti p130cas, by Cell Signaling Technology (1 mentions) Anti phospho p130cas, by Cell Signaling Technology (1 mentions) Anti erbb2, by Cell Signaling Technology (1 mentions) Anti gfp, by Abcam (1 mentions)

5 protocols using anti nrp 1

Immunofluorescence Staining of Endothelial Cells

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After been fixed with 4% (w/v) paraformaldehyde for 30 min, cells were permeabilized with 0.1% (v/v) Triton X-100 in PBS for 5 min. A total of 10% (v/v) goat serum was added, and the cells were incubated for 30 min to block unspecific antibody binding. Next, the cells were incubated with the following primary antibodies overnight at 4°C: anti-CD31 (Santa Cruz Biotechnology), anti-CD144 (eBioscience), anti–NRP-1 (Santa Cruz Biotechnology), and anti–α-SMA, (Chemicon). Then, the cells were washed with PBS three times and incubated with Alexa Fluor 488– or Alexa Fluor 565–conjugated secondary antibodies (Molecular Probe). The stained cultures were counterstaining with 4′,6-diamidino-2-phenylindole (DAPI) (DAKO). Confocal images were acquired with an Olympus FV1000MPE confocal microscope using an objective lens (UPlanSApo 60XW, NA 1.20, Olympus). Z-stack images were taken with 10-μm intervals. Images were analyzed using FV10-ASW 3.0 Viewer software (Olympus) and processed in ImageJ software.

Gil C.H., Chakraborty D., Vieira C.P., Prasain N., Calzi S.L., Fortmann S.D., Hu P., Banno K., Jamal M., Huang C., Sielski M.S., Lin Y., Huang X., Dupont M.D., Floyd J.L., Prasad R., Longhini A.L., McGill T.J., Chung H.M., Murphy M.P., Kotton D.N., Boulton M.E., Yoder M.C, & Grant M.B. (2022). Specific mesoderm subset derived from human pluripotent stem cells ameliorates microvascular pathology in type 2 diabetic mice. Science Advances, 8(9), eabm5559.

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Western Blot Protein Quantification

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For protein quantification, total cell extracts were fractioned by electrophoresis on SDS-PAGE followed by transferring onto a PVDF membrane. Western blotting was performed with the following primary antibodies: anti-beta Catenin (ZYMED, 13-8400), anti-NRP1 (Santa Cruz Biotechnology, sc-7239), and anti-beta-actin (Cell Signaling, 3700S).

Xu X., Zhang Y., Jasper J., Lykken E., Alexander P.B., Markowitz G.J., McDonnell D.P., Li Q.J, & Wang X.F. (2016). MiR-148a functions to suppress metastasis and serves as a prognostic indicator in triple-negative breast cancer. Oncotarget, 7(15), 20381-20394.

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Immunofluorescent Characterization of ECFCs

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ECFCs were fixed with 4% (w/v) paraformaldehyde for 30 min and permeabilized with 0.1% (v/v) Triton X-100 in PBS for 5 min. After blocking with 10% (v/v) goat serum for 30 min, cells were incubated with primary following antibodies; anti-CD31 (Santa Cruz), anti-CD144 (Ebioscience), anti-NRP-1 (Santa Cruz) and anti-a-SMA, (Chemicon) overnight at 4 °C. Cells were washed with PBS, then incubated with secondary antibodies conjugated with Alexa-488 or Alexa-565 (Molecular Probe) and visualized by confocal microscopy after counterstaining with 2 g/ml DAPI (Sigma-Aldrich). The confocal images were obtained with an Olympus FV1000 mpE confocal microscope using as an Olympus uplanSApo 60xW/1.2NA/eus objective. All the images were taken as Z-stacks with individual 10-μm thick sections at room temperature and images were analyzed using FV10-ASW 3.0 Viewer.

Prasain N., Lee M.R., Vemula S., Meador J.L., Yoshimoto M., Ferkowicz M.J., Fett A., Gupta M., Rapp B.M., Saadatzadeh M.R., Ginsberg M., Elemento O., Lee Y., Voytik-Harbin S.L., Chung H.M., Hong K.S., Reid E., O'Neill C.L., Medina R.J., Stitt A.W., Murphy M.P., Rafii S., Broxmeyer H.E, & Yoder M.C. (2014). Differentiation of human pluripotent stem cells to cells similar to cord-blood endothelial colony–forming cells. Nature biotechnology, 32(11), 1151-1157.

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SDS-PAGE Immunoblotting for Protein Detection

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SDS-PAGE electrophoresis and transfer of separated polypeptides to nitrocellulose membranes (GE Healthcare Europe, Milano, Italy) was performed by standard techniques using 60 μg of proteins per sample in 7-12% gradient SDS-polyacrylamide gels. Detection of the different polypeptides under study was performed using the following primary antibodies: anti-PDGFRα (polyclonal rabbit antibody C20) and anti-NRP-1 (monoclonal mouse antibody A12) from Santa Cruz Biotechnology (Santa Cruz, CA); anti-p130Cas (monoclonal rabbit antibody E1L9H) and anti-phospho-p130Cas (rabbit polyclonal antibody Tyr410) from Cell Signaling Technology (Danvers, MA); anti-β-tubulin rabbit polyclonal antibody H-235 from Santa Cruz Biotechnology; EMT markers, from the Cell Signaling EMT antibody sampler kit.

Ruffini F., Levati L., Graziani G., Caporali S., Atzori M.G., D’Atri S, & Lacal P.M. (2017). Platelet-derived growth factor-C promotes human melanoma aggressiveness through activation of neuropilin-1. Oncotarget, 8(40), 66833-66848.

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Immunohistochemical and X-Gal Analysis of Cardiac Tissues

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Samples were harvested, fixed overnight in 2–4% paraformaldehyde and dehydrated through an ethanol series. All samples were paraffin embedded and sectioned. Antibodies used for immunofluorescence were anti-ErbB2 (Cell Signaling, 1:50), anti-phoshpo-ErbB2 Y1248 (Cell Signaling, 1:50), anti-Nrp1 (Santa Cruz, 1:25), anti-GFP (Abcam, 1:250), and anti-eNOS (BD Bioscieznces, 1:250). Hematoxylin and eosin staining was completed using a standard protocol.
For whole-mount X-gal staining, adult and embryonic hearts were isolated and fixed with 2% paraformaldehyde in PBS for 20 min at 4 °C. Following fixation, the samples were washed twice for 10 min in PBS at 4 °C. Embryos were stained in with PBS containing 1 mg ml⁻¹ X-gal, 5 mM K₃Fe(CN)₆, 5 mM K₄Fe(CN)₆, 2 mM MgCl₂, 0.01% NP-40, 0.01% sodium deoxycholate and incubated overnight at 37 °C.

Aghajanian H., Cho Y.K., Manderfield L.J., Herling M.R., Gupta M., Ho V.C., Li L., Degenhardt K., Aharonov A., Tzahor E, & Epstein J.A. (2016). Coronary vasculature patterning requires a novel endothelial ErbB2 holoreceptor. Nature Communications, 7, 12038.

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