Goat anti nrp1

Manufactured by R&D Systems

Sourced in Germany

Goat anti-NRP1 is a polyclonal antibody that recognizes the NRP1 (Neuropilin-1) protein. NRP1 is a transmembrane glycoprotein that functions as a cell surface receptor for several ligands, including VEGF, semaphorins, and other extracellular matrix proteins. This antibody can be used for the detection and study of NRP1 in various biological applications.

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Lab products found in correlation

Lsm 700, by Zeiss (4 mentions) Goat anti nrp2, by R&D Systems (3 mentions) Cm1850 cryostat, by Leica (2 mentions) Chicken anti gfp, by Abcam (2 mentions) Prolong gold antifade reagent, by Thermo Fisher Scientific (2 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (2 mentions) Alexa fluor 488 donkey anti goat, by Thermo Fisher Scientific (1 mentions) Guinea pig anti vglut2, by Synaptic Systems (1 mentions) Phospho creb, by Cell Signaling Technology (1 mentions) Af566, by R&D Systems (1 mentions) Rabbit anti synaptophysin, by Cell Signaling Technology (1 mentions) Rabbit anti phox2b, by Abcam (1 mentions) Rabbit anti ntrk1, by Merck Group (1 mentions) Rabbit anti cleaved caspase 3, by Cell Signaling Technology (1 mentions) Mouse anti s100β, by Abcam (1 mentions) Mouse anti α smooth muscle actin α sma, by Abcam (1 mentions) Alexa 647 conjugated secondary antibody, by Jackson ImmunoResearch (1 mentions) Antigen retrieval solution, by BioGenex (1 mentions) Goat anti sox10, by Santa Cruz Biotechnology (1 mentions) Prolong gold antifade reagent with dapi, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

Anti-NRP1 (4 citations) Goat anti-mouse Nrp1 (2 citations) Polyclonal goat anti-mouse Nrp1 and mouse CD22-Fc fusion proteins (2 citations) Goat anti-rat/mouse Nrp1 (2 citations)

10 protocols using goat anti nrp1

Sublethal and Lethal Dose TBI Therapy

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For sublethal TBI, adult C57BL/6J mice were irradiated with 500 cGy TBI using a Cesium-137 irradiator. Mice were dosed via intravenous injection (IV) every other day for up to 10 days with 10 µg goat anti-NRP1 (R&D Systems, Minneapolis, MN), or goat IgG control (R&D Systems, Minneapolis, MN), beginning at +24 h of post-TBI.
For lethal dose TBI, adult C57BL/6J mice were irradiated with 800 cGy TBI, which is lethal at our institution for approximately 50% of C57BL/6J mice by day +30 (LD50/30). Mice were dosed IV with either 10 µg goat anti-NRP1 (R&D Systems, Minneapolis, MN) or goat IgG control (R&D Systems, Minneapolis, MN), beginning at +24 h of post-TBI and every other day through day +21.
For evaluation of chemotherapy effects, adult C57BL/6J mice were treated with 5 fluorouracil (5FU), 250 mg/kg IV × 1, and then administered 10 μg anti-NRP1 or goat IgG control every other day IV through day +10.

Termini C.M., Pang A., Fang T., Roos M., Chang V.Y., Zhang Y., Setiawan N.J., Signaevskaia L., Li M., Kim M.M., Tabibi O., Lin P.K., Sasine J.P., Chatterjee A., Murali R., Himburg H.A, & Chute J.P. (2021). Neuropilin 1 regulates bone marrow vascular regeneration and hematopoietic reconstitution. Nature Communications, 12, 6990.

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Whole-mount Immunostaining of Brain Tissue

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Samples were processed according to the iDISCO protocol (Renier et al., 2014 (link)). Primary antibodies were goat anti-Nrp1 (#AF566; R&D Systems) and guinea pig anti-VGLUT2 (#135 404; Synaptic Systems, Göttingen, Germany). Secondary antibodies were donkey anti-goat Alexa Fluor 488 (#A11055; Thermo Fisher Scientific) and donkey anti-guinea pig RRX (#706-295-148; Jackson ImmunoResearch Laboratories).

Zapiec B., Bressel O.C., Khan M., Walz A, & Mombaerts P. (2016). Neuropilin-1 and the Positions of Glomeruli in the Mouse Olfactory Bulb. eNeuro, 3(5), ENEURO.0123-16.2016.

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Immunohistochemistry Antibody Panel for Neuronal Markers

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The antibodies used are: mouse anti-Meis1 (Millipore, 05–779, 1/100 for immunoblotting, Germany), goat anti-Meis1 (Santa Cruz, sc-10599, 1/100 for immunochemistry, Dallas, Texas, USA), sheep anti-TH (Thermo Scientific, PA1-4679, 1/2000, Waltham, MA, USA), rabbit anti-PGP9.5 (Millipore, AB1761, 1/500, Germany), goat anti-Nrp-1 (R&D Systems, AF566, 1/250, Minneapolis, USA), rabbit anti-Synaptophysin (Cell Signaling Technology, 5461, 1/250, Danvers, MA, USA), rabbit anti-Phox2b (kindly provided by JF Brunet, 1/250, or Abcam ab183741, 1/250, UK), goat anti-c-Ret (R&D Systems, AF482, 1/20000, Minneapolis, USA), rabbit anti-Ntrk1 (Millipore, 06–574, 1/250, Germany), rabbit anti-Cleaved Caspase-3 (Cell Signaling Technology, 9661, 1/250, Danvers, MA, USA), rabbit anti-Clathrin Heavy Chain 1 (Thermo Scientific, PA5-17347, 1/200, Waltham, MA, USA), Phospho-CREB (Cell Signaling, 87G3, 1/200, Germany), Rab5 (Abcam, ab18211, 1/200, UK), Sox2 (Santa Cruz, sc-17320, 1/250, Dallas, Texas, USA).

Bouilloux F., Thireau J., Ventéo S., Farah C., Karam S., Dauvilliers Y., Valmier J., Copeland N.G., Jenkins N.A., Richard S, & Marmigère F. (2016). Loss of the transcription factor Meis1 prevents sympathetic neurons target-field innervation and increases susceptibility to sudden cardiac death. eLife, 5, e11627.

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Immunostaining of Enteric Neurons and Glia

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Segments of distal colon from 7- and 36-day-old rats were fixed in 0.1 M phosphate-buffered saline (PBS) containing 4% paraformaldehyde (PFA) at room temperature (RT) for 3 h. Whole mounts of longitudinal muscle and myenteric plexus (LMMP) were obtained by microdissection and were permeabilized with PBS containing 10% BSA, 0.5% Triton X-100 and 0.01% sodium azide for 2 h at RT. Tissues were then incubated with the following primary antibodies: rabbit anti-Sema3A (Santa Cruz, 1/200), goat anti-NRP1 (R&D System, 1/33), mouse anti-α smooth muscle actin (αSMA, Abcam, 1/500), mouse or rabbit anti-TuJ1 (Sigma or Abcam respectively, 1/1,000), mouse anti-S100β (Abcam, 1/500) and mouse or rabbit anti-HuC/D (HU; Molecular Probes or Santa Cruz respectively, 1/500) diluted in PBS containing 3% BSA, 0.25% Triton X-100 and 0.01% sodium azide for 16 h at RT. After washing, tissues were incubated for 2 h at RT with the appropriate Alexa 488-, Cy3-, Cy5- or Alexa 647-conjugated secondary antibodies (Jackson ImmunoResearch), and then incubated for 5 min in DAPI (ThermoFisher Scientific, 1/5,000 and mounted with ProLong Gold Antifade Reagents (ThermoFisher Scientific).

Gonzales J., Le Berre-Scoul C., Dariel A., Bréhéret P., Neunlist M, & Boudin H. (2020). Semaphorin 3A controls enteric neuron connectivity and is inversely associated with synapsin 1 expression in Hirschsprung disease. Scientific Reports, 10, 15119.

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Immunofluorescence Analysis of Embryonic Markers

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X-gal staining for detection of β-galactosidase and counterstaining with Nuclear Fast Red were conducted as described previously (Anderson et al., 2004 (link)). For section immunofluorescence, embryos were sectioned, dewaxed, boiled in antigen retrieval solution (Biogenex), and blocked in PBS containing 10% sheep serum and 0.1% Triton X-100. The following primary antibodies were used at 1:100 dilutions in blocking serum: goat anti-ECE1 (R&D AF1784), goat anti-Nrp1 (R&D AF566), goat-anti Nrp2 (R&D AF2215), goat anti-connexin40 (Santa Cruz sc-20466), mouse anti-COUP-TFII (Perseus Proteomics PP-H7147-00), and chicken anti-β-galactosidase (Abcam ab9361). The following secondary antibodies were used: rabbit anti-goat 594 (Invitrogen A11080), rabbit anti-chicken 488 (Jackson ImmunoResearch 303-545-003), and rabbit anti-mouse 594 (Invitrogen A11062). Slides were mounted and photographed as described previously (Rojas et al., 2009 (link)).

Robinson A.S., Materna S.C., Barnes R.M., De Val S., Xu S.M, & Black B.L. (2014). An arterial-specific enhancer of the human Endothelin-converting enzyme 1 (ECE1) gene is synergistically activated by Sox17, FoxC2, and Etv2. Developmental biology, 395(2), 379-389.

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Immunolabeling of Developing Neurons

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Embryos were fixed in 4% formaldehyde in PBS. All sections were cut at 18 µm on a CM1850 cryostat (Leica) and air-dried for 60 min before immunolabelling. For immunolabelling, cryosections were blocked in 0.2% BSA and 0.5% Triton X-100 in PBS, and stained using the indicated primary antibodies. GFP⁺ NCCs grown on fibronectin-coated slides were fixed in 4% formaldehyde in PBS for 10 min, blocked in 10% DAKO block, 0.5% Triton X-100 in PBS and stained with the indicated primary antibodies. Antibodies used were: rabbit anti-TH (Millipore), 1:300; goat anti-NRP1 (R&D), 1:500; rabbit anti-NRP1 (Abcam), 1:500; rabbit anti-NRP2 (CST), 1:500; mouse anti-TUJ1 (Sigma Aldrich), 1:750; chicken anti-GFP, 1:1000 (Abcam); goat anti-SOX10 (Santa Cruz), 1:200; and rabbit anti-VAChT (Sy Sy), 1:1000. Cryosections and cells were mounted in Prolong Gold antifade reagent with DAPI (Molecular Probes). Confocal images were acquired on a LSM 700 (Zeiss) system. All images were prepared with Adobe Photoshop (Adobe).

Lumb R., Tata M., Xu X., Joyce A., Marchant C., Harvey N., Ruhrberg C, & Schwarz Q. (2018). Neuropilins guide preganglionic sympathetic axons and chromaffin cell precursors to establish the adrenal medulla. Development (Cambridge, England), 145(21), dev162552.

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Isolation and Analysis of Immune Cells from Spleen, Tumor, and Lymph Nodes

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The spleen, tumor or tumor-draining lymph node (TDLN) cell isolation procedure was based on our previously described protocol55 (link). Tissues were cut into fragments and then incubated in 1 mg/ml collagenase III (Gibco-Invitrogen) in RPMI 1640 at 37 °C for 15 min. Flow cytometry staining was performed using fluorescence-conjugated anti–mouse antibodies targeting CD4, CD8a, CD69, CD25, IFN-γ, IL17A, CD11b and CD11c (BD Pharmingen). Cells were also incubated with goat anti-Nrp-1 (R&D Systems) and rat anti-TGF-β1 (Biolegend) followed by an Alex Fluor 647-conjugated anti-goat antibody (BD Pharmingen) or Alex Fluor 488-conjugated anti-rat antibody (BD Pharmingen). Intracellular staining was performed using a Foxp3 staining kit and fixation/permeabilization buffer (eBioscience). Samples were obtained using a FACSCalibur equipped with Cell Quest software (BD Biosciences). The cells were gated for lymphocytes based on the forward and side scatter profiles. All data were analyzed using Tree Star FlowJo software.

Wang S., Gao X., Shen G., Wang W., Li J., Zhao J., Wei Y.Q, & Edwards C.K. (2016). Interleukin-10 deficiency impairs regulatory T cell-derived neuropilin-1 functions and promotes Th1 and Th17 immunity. Scientific Reports, 6, 24249.

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Immunolabeling of Embryonic Tissue Sections

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Embryos were fixed in 4% paraformaldehyde in PBS. All sections were cut at a thickness of 14 μm on a CM1850 cryostat (Leica, North Ryde, NSW, Australia) and air-dried for 60 min before staining. For immunolabelling, cryosections or whole mount embryos were blocked in PBS containing 0.2% BSA and 0.5% Triton X-100, and stained with the indicated primary antibodies: rabbit anti-Sox9 1:1000; rabbit tyrosine hydoxylase 1:300 (both Millipore, Batswater, VIC, Australia); goat anti-Nrp1 1:500; rabbit anti-Nrp2 1:500 (both R&D, Minneapolis, MN, USA); rabbit anti-p75-NTR 1:200 (Epitomics, Burlingame, CA, USA); mouse anti β-tubulin isotype III (Sigma-Aldrich, Sydney, NSW, Australia) 1:750; and chicken anti-GFP 1:1000 (Abcam, Melbourne, VIC, Australia). Cyrosections were mounted in Prolong Gold Antifade Reagent containing DAPI (Molecular Probes, Mulgrave, VIC, Australia). Confocal images were acquired under a confocal microscope (LSM 700; Zeiss, Jena, Germany). All images were prepared with Photoshop software (Adobe, San Jose, CA, USA).

Lumb R., Wiszniak S., Kabbara S., Scherer M., Harvey N, & Schwarz Q. (2014). Neuropilins define distinct populations of neural crest cells. Neural Development, 9, 24.

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Immunofluorescence Staining for Cell Markers

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Single cells were fixed at 4 % PFA (in 1X PBS, pH 7.4) for 10 min at room temperature (RT). Brain slices and dissociated single cells were washed in 1X PBS (pH 7.4) with 0.2 % Tween 20, followed by blocking (in 4 % BSA in 1X PBS/0.2 % Tween 20) for 2h at RT and incubation with the primary antibody at 4 °C, overnight. After washing, the secondary antibody was applied for 2h at RT. The nuclei were stained with DAPI (1 ng/ml in H2O) for 15 min at RT. As primary antibodies were used: goat anti Nrp1 (R&D, 1:100), goat anti Nrp2 (R&D, 1:100), rabbit anti Calbindin (Swann, 1:600) and Placental alkaline phosphatase (PLAP) (ABD-serotec, 1:100). As secondary antibodies were used: donkey anti rabbit FP 647H (Interchim, 1:1000) and donkey anti goat Alexa555 (Molecular probes, 1:1000). In the case of the co-localization, after fixation, one additional washing step was performed with warm PBS (65 °C) to eliminate any intrinsic alkaline phosphatase present in the cells. At the stripe assay, in order to visualize the Human Sema3C-Fc recombinant chimera, bound to the stripes, anti-human conjugated to Alexa 488 (Invitrogen, 30 μg/ml), was applied after fixation, following the same procedure as with a secondary antibody.

Aiello K., & Bolz J. (2021). Semaphorin 3C attracts MGE-derived cortical interneurons in the deep migratory stream guiding them into the developing neocortex.

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Immunostaining Protocol for Single Cells and Brain Slices

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Single cells were xed at 4% PFA (in 1X PBS, pH 7.4) for 10 min at room temperature (RT). Brain slices and dissociated single cells were washed in 1X PBS (pH 7.4) with 0.2% Tween 20, followed by blocking (in 4% BSA in 1X PBS/0.2% Tween 20) for 2h at RT and incubation with the primary antibody at 4°C, overnight. After washing, the secondary antibody was applied for 2h at RT. The nuclei were stained with DAPI (1 ng/ml in H2O) for 15 min at RT. As primary antibodies were used: goat anti Nrp1 (R&D, 1:100), goat anti Nrp2 (R&D, 1:100), rabbit anti Calbindin (Swann, 1:600) and Placental alkaline phosphatase (PLAP) (ABD-serotec, 1:100). As secondary antibodies were used: donkey anti rabbit FP 647H (Interchim, 1:1000) and donkey anti goat Alexa555 (Molecular probes, 1:1000). In the case of the co-localization, after xation, one additional washing step was performed with warm PBS (65°C) to eliminate any intrinsic alkaline phosphatase present in the cells. At the stripe assay, in order to visualize the Human Sema3C-Fc recombinant chimera, bound to the stripes, anti-human conjugated to Alexa 488 (Invitrogen, 30 µg/ml), was applied after xation, following the same procedure as with a secondary antibody.

Aiello K., & Bolz J. (2022). Semaphorin 3C attracts MGE-derived cortical interneurons in the deep migratory stream guiding them into the developing neocortex.

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