Goat anti notch1

Manufactured by Santa Cruz Biotechnology

Sourced in United Kingdom

Goat anti-Notch1 is a primary antibody that binds to the Notch1 protein. Notch1 is a transmembrane receptor that plays a critical role in cell-cell signaling and cell fate determination during development and adult tissue homeostasis. This antibody can be used to detect and study the expression and localization of Notch1 in various biological samples.

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Goat polyclonal anti-Notch1 Anti-Notch1 Notch1

3 protocols using goat anti notch1

Lung Tissue Protein Expression Analysis

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Lung tissue (200 mg) was homogenized in RIPA lysis buffer (100 ml PBS 1x pH 7.2, 1% NP-40, 0.5% sodium deoxycholate, 0.1% SDS, supplemented with protease inhibitors). Total protein was quantified using Bradford assay (Bio-Rad, Hercules, CA). Samples were subjected to SDS-PAGE electrophoresis, using a 15% acrylamide gel for CC10 and 10% acrylamide for Notch1. Seventy five μg of protein were loaded for CC10 and 50 μg for Notch1. They were transferred to PVDF membranes and blocked with 5% nonfat milk. The following primary antibodies were used: mouse anti-Muc5ac (1:2000, sc-21701, Santa Cruz Biotechnology), goat anti-Muc5b (1:2000, sc-135508, Santa Cruz Biotechnology), rabbit anti-CC10 (1:4000, ab40873, Abcam, UK), goat anti-Notch1 (1:2000, sc-6015, Santa Cruz Biotechnology), and Anti-Actin I-19 (1:2000, sc-1616, Santa Cruz Biotechnology, USA). Secondary antibodies conjugated to horseradish peroxidase (Santa Cruz Biotechnology) were used. Brain and stomach were used as positive controls. Water without antibodies was used as negative control. Proteins were detected by Pierce, ECL Western Blotting Substrate (Pierce Biotechnology, Rockford, IL). Images were film-captured. Density of band was measured using Fiji software (ImageJ 2.0.0-rc-54/1.51h) and protein amounts were normalized with the actin signal.

Méndez A., Rojas D.A., Ponce C.A., Bustamante R., Beltrán C.J., Toledo J., García-Angulo V.A., Henriquez M, & Vargas S.L. (2019). Primary infection by Pneumocystis induces Notch-independent Clara cell mucin production in rat distal airways. PLoS ONE, 14(6), e0217684.

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Immunofluorescence Analysis of Angiogenic Markers

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For cytospins, C6 glioma cells in endothelial differentiation medium were seeded on coverslips and cultured for 24 h in hypoxia. Tumors were collected and then divided for frozen section at 5μm thickness. Sections were fixed in 4% paraformaldehyde and blocked with 10% goat serum. The primary antibodies used in this study were as follows: rabbit anti-von Willebrand factor (vWF) (Millipore, Bedford, MA), rabbit anti-CD34, mouse anti-CD31, mouse anti-Flk1 (Santa Cruz Biotechnology Inc., Santa Cruz, CA), and goat anti-Notch1 (Santa Cruz Biotechnology Inc., Santa Cruz, CA). The secondary antibodies used were as follows (all from Invitrogen, Grand Island, NY): Alexa Fluor 555-donkey anti-rabbit IgG, Alexa Fluor 568-rabbit anti-mouse IgG, Alexa Fluor 568-rabbit anti-goat IgG, Alexa Fluor 647-rabbit anti-mouse IgG, Alexa Fluor 647-donkey anti-rabbit IgG. Antibodies were diluted in antibody diluent (Beyotime, Jiangsu, China), and incubations were done at room temperature. The images were captured by confocal laser scanning microscopy (Leica, Heerbrugg, Switzerland), and the obtained images were processed by Adobe Photoshop 7.0 (Adobe System Inc., San Jose, CA).

Chen X., Fang J., Wang S., Liu H., Du X., Chen J., Li X., Yang Y., Zhang B, & Zhang W. (2014). A new mosaic pattern in glioma vascularization: exogenous endothelial progenitor cells integrating into the vessels containing tumor-derived endothelial cells. Oncotarget, 5(7), 1955-1968.

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Quantitative Protein Analysis of DRG Neurons

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Total proteins from cultured DRG neurons were isolated on DIV6 according to published protocols [25 (link), 27 (link)]. Total protein from DRG, sciatic nerve and foot pad tissues of db/db and db/m mice were extracted using the same methods. In vitro samples from 4 individual microfluidic chambers were pooled for one Western blot. The protein concentration was measured using a bicinchoninic acid protein assay kit (Thermo Fisher Scientific). Western blot was performed according to previously described methods [25 (link), 27 (link)]. Briefly, equal amounts of proteins were loaded. Primary antibodies were rabbit anti-ADAM10 (a disintegrin and metalloproteinase domain-containing protein 10, 1:1000, Abcam, Cambridge, UK), rabbit anti-DCX (doublecortin, 1:500, Abcam), rabbit anti-c-MET (tyrosine-protein kinase met, 1:500, Abcam), rabbit anti-FOXP2 (1:1000, Abcam), goat anti-NOTCH1 (1:1000, Santa Cruz), rabbit anti-ROCK1 (Rho associated coiled-coil containing protein kinase 1, 1:500, Abcam), rabbit anti-SYNJ1 (Synaptojanin 1, 1:1000, Sigma-Aldrich), rabbit anti-VAMP2 (vesicle-associated membrane protein 2, 1:1000, Cell Signaling Technology, Danvers, MA, USA), goat anti-VAT1 (1:1000, Santa Cruz), and mouse anti-β-actin (1:10000; Abcam). The optical density of protein bands was measured and calculated by means of Fluorchem E instrument (ProteinSimple, San Jose, CA, USA).

Jia L., Chopp M., Wang L., Lu X., Zhang Y., Szalad A, & Zhang Z.G. (2018). MiR-34a Regulates Axonal Growth of Dorsal Root Ganglia Neurons by Targeting FOXP2 and VAT1 in Postnatal and Adult Mouse. Molecular neurobiology, 55(12), 9089-9099.

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