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P75NTR is a receptor protein that binds to and interacts with the p75 neurotrophin receptor. It plays a role in neuronal signaling and function. The core function of P75NTR is to facilitate cellular responses to neurotrophins, which are proteins that promote the survival and differentiation of neurons.

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Lab products found in correlation

Sortilin, by BD (2 mentions) β actin, by Merck Group (2 mentions) Ykl 40, by Abcam (1 mentions) 3 3 diaminobenzidine (dab), by Agilent Technologies (1 mentions) Envision system hrp, by Agilent Technologies (1 mentions) Mms 435p, by Merck Group (1 mentions) P mapk, by Cell Signaling Technology (1 mentions) P akt, by Cell Signaling Technology (1 mentions) P jnk, by Cell Signaling Technology (1 mentions) Odyssey infrared imaging system, by LI COR (1 mentions) P creb, by Cell Signaling Technology (1 mentions) Pro ngf, by Merck Group (1 mentions) Western blotting reagents, by Cytiva (1 mentions) Phospho sapk jnk, by Cell Signaling Technology (1 mentions) Fitc annexin 5 apoptosis detection kit, by BD (1 mentions) Horseradish peroxidase hrp conjugated secondary antibody, by Santa Cruz Biotechnology (1 mentions) Alexa fluor 488 conjugated secondary antibody, by Thermo Fisher Scientific (1 mentions) Brain derived neurotrophic factor (bdnf), by Santa Cruz Biotechnology (1 mentions) Nerve growth factor (ngf), by Santa Cruz Biotechnology (1 mentions) Probdnf antibody, by Alomone (1 mentions)

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Anti-p75NTR (5 citations)

6 protocols using p75ntr

1

Immunohistochemical Analysis of Tumor Markers

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Animals were sacrificed after four weeks and tumors were removed. Hematoxylin–eosin (H&E) staining and immunohistochemistry were performed on 4μm serial coronal sections from paraffin-embedded tumors. Tissue sections were prepared as previously described [62 (link)]. Tissue sections were incubated with antibodies against human Ki67 (1/50, Dako), CD31(1/30, Histonova), YKL-40 (1/1000, Abcam), TrkB (1/200, R&D), p75NTR (1/300, Santa Cruz), sortilin (1/1000, BD bioscience) and Oct4 (1/200) according to the manufacturer's instructions (Boster Bioengineering Company Limited). Anti-rabbit (1/1000) and anti-mouse (1/1000) immunoglobulins HRP EnVision™+ system (Dakocytomation, Glostrup) and DAB (DakoCytomation) were used for the staining revelation.
Pinet S., Bessette B., Vedrenne N., Lacroix A., Richard L., Jauberteau M.O., Battu S, & Lalloué F. (2016). TrkB-containing exosomes promote the transfer of glioblastoma aggressiveness to YKL-40-inactivated glioblastoma cells. Oncotarget, 7(31), 50349-50364.
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2

Western Blot Analysis of Neuronal Signaling

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Brain homogenates were resolved by SDS–polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to activated PVDF membranes. The membranes were blocked with 5% bovine serum albumin in tris-buffered saline/Tween 20 (TBST). The following primary antibodies were incubated overnight at 4°C: TrkA (1:1000; Cell Signaling, 2505), p75NTR (1:1000; Santa Cruz Biotechnology, sc-271708), pAkt (1:1000; Cell Signaling, 4060), Akt (1:1000; Cell Signaling, 9272), pCREB (1:1500; Cell Signaling, 9198), CREB (1:1500; Cell Signaling, 9104), pMAPK (1:2000; Cell Signaling, 9101), MAPK (1:2000; Cell Signaling, 9102), pJNK (1:1000; Cell Signaling, 9251), JNK (1:1000; Cell Signaling, 9252), βtubIII (1:5000; Covance, MMS-435P), and β-actin (1:5000; Sigma-Aldrich, A5316). Membranes were washed in TBST, incubated with corresponding IRDye secondary antibodies at 1:15,000 for 1 hour at RT (LI-COR Biosciences), and imaged with an Odyssey infrared imaging system (LI-COR Biosciences). Densitometry analysis was performed using ImageStudio software.
Xhima K., Markham-Coultes K., Nedev H., Heinen S., Saragovi H.U., Hynynen K, & Aubert I. (2020). Focused ultrasound delivery of a selective TrkA agonist rescues cholinergic function in a mouse model of Alzheimer’s disease. Science Advances, 6(4), eaax6646.
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3

Antibody Sourcing for Western Blot Analysis

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Antibodies against beta-actin, LAMP1, P0, p62 and p75NTR were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). Anti-phospho-ERK, anti-phospho-AKT and anti-phsopho-p70 S6 kinase antibodies were obtained from Cell Signaling Lab (Beverly, MA). Antibodies against MBP and S100 were obtained from Abcam and Chemicon, respectively. Antibodies against ATG7 were obtained from Sigma (St. Louis, MO) and Cell Signaling Lab. Antibody against LC3B was purchased from Invitrogen (Carlsbad, CA). All other undesignated reagents were purchased from Sigma.
Jang S.Y., Shin Y.K., Park S.Y., Park J.Y., Rha S.H., Kim J.K., Lee H.J, & Park H.T. (2015). Autophagy Is Involved in the Reduction of Myelinating Schwann Cell Cytoplasm during Myelin Maturation of the Peripheral Nerve. PLoS ONE, 10(1), e0116624.
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4

Nanoparticle Effects on Neurotrophin Signaling

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Commercial grade titanium dioxide nanoparticles (TiO2-NP) and titanium(IV) oxide fine particles (TiO2-FP) were kindly provided by the National Institute for Occupational Safety and Health (NIOSH). MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] cell proliferation assay kits were purchased from Cayman Chemical (Ann Arbor, MI); FITC Annexin-V Apoptosis Detection kits from BD Biosciences (San Jose, CA); and RNeasy kits for total RNA isolation from Qiagen (Valencia, CA). Primers for human NGF, BDNF, TrkA, TrkB and p75NTR were purchased from SABiosciences (Valencia, CA) and primers for the β2 microglobulin (B2M) housekeeping gene from RealTimePrimers (Elkins Park, PA).
The following primary antibodies, secondary antibodies, and isotype controls were used for immunostaining: NGF, BDNF, p75NTR, GAPDH and JNK (Santa Cruz Biotechnology, Santa Cruz, CA); phospho-SAPK/JNK (Cell Signaling, Danvers, MA); pro-NGF (Millipore, Billerica, MA); TrkA and TrkB (R&D Systems, Minneapolis, MN); AlexaFluor 488-conjugated secondary antibodies (Invitrogen, Carlsbad, CA); and horseradish peroxidase (HRP)-conjugated secondary antibodies (Santa Cruz Biotechnology). Western blotting reagents were purchased from Amersham Life Science (Buckinghamshire, United Kingdom). Interleukin 1 alpha (IL-1α) and IL-1 receptor antagonist (IL1-ra) were purchased from Santa Cruz Biotechnology.
Chakraborty S., Castranova V., Perez M.K, & Piedimonte G. (2017). Nanoparticles-Induced Apoptosis of Human Airway Epithelium is mediated by ProNGF/p75NTR Signaling. Journal of toxicology and environmental health. Part A, 80(1), 53-68.
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5

Protein Signaling Pathway Analysis via Western Blotting

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Western blotting was performed with specific antibodies against phospho-p38MAPK-T180/Y182 (#4511), or phospho-AKT-T308 (#2965) from Cell Signaling, or phospho-ERK1(T202/Y204)/ERK2(T185/Y187) (R&D AF1018) and ERK1/2 (AF1576) were obtained from R&D. Antibodies against whole form of p38MAPK (#8690), and AKT (#4691) were from Cell Signaling. p75NTR (Santa Cruz, sc-8317) or (Abcam, ab52987). Pro-BDNF antibody was purchased from Alomone (Cat#: ANT-006). Trk-B (#610101) and sortilin (#612101) were from BD company. As loading control β-actin (Sigma-Aldrich A5441) was used. Functional assays were realized with specific antagonistic antibody to block p75NTR (Upstate, clone ME 20.4, cat# 05-446) used at 15 ng/mL.
In some experiments, we used a broad-spectrum inhibitor of matrix metalloproteinases (MMPs) GM6001 (Calbiochem, CAS142880-36-2) (20 μM) to inhibit the cleavage of pro-BNDF in mature BDNF [21 (link)]. In all cases, GM6001 was added to culture media 30 minutes before pro-BDNF treatment.
De la Cruz-Morcillo M.A., Berger J., Sánchez-Prieto R., Saada S., Naves T., Guillaudeau A., Perraud A., Sindou P., Lacroix A., Descazeaud A., Lalloué F, & Jauberteau M.O. (2016). p75 neurotrophin receptor and pro-BDNF promote cell survival and migration in clear cell renal cell carcinoma. Oncotarget, 7(23), 34480-34497.
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6

Silencing p75NTR in U937 Cells

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U937 cells were electroporated with control CRISPR-Cas9 plasmids or CRISPR-Cas9 plasmids containing gRNA that targeted p75NTR (Santa Cruz Biotechnology). The Gene Pulser MXcell electroporation system (Bio-Rad Laboratories, Hercules, CA, USA) was used at 210 V and 960 μF. The cells were then cultured in Iscove’s Modified Dulbecco’s Medium (IMDM) (Invitrogen, Carlsbad, CA, USA) with 10% fetal bovine serum (Invitrogen) and 0.3 μg/mL puromycin (Sigma-Aldrich, St. Louis, MO, USA). Cells that survived puromycin selection were isolated and validated by Western blot analysis.
Yu H.C., Huang H.B., Huang Tseng H.Y, & Lu M.C. (2022). Brain-Derived Neurotrophic Factor Suppressed Proinflammatory Cytokines Secretion and Enhanced MicroRNA(miR)-3168 Expression in Macrophages. International Journal of Molecular Sciences, 23(1), 570.
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