Rabbit anti trkb

Manufactured by Cell Signaling Technology

Sourced in United States

Rabbit anti-TrkB is a primary antibody that specifically recognizes the TrkB receptor. TrkB is a receptor tyrosine kinase that binds to the neurotrophin brain-derived neurotrophic factor (BDNF) and mediates BDNF signaling in the nervous system.

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Lab products found in correlation

Mouse anti neun, by Merck Group (2 mentions) Mouse anti α tubulin, by Merck Group (2 mentions) Mouse anti flag, by Merck Group (2 mentions) Alexa fluor 594 conjugated secondary antibody, by Thermo Fisher Scientific (1 mentions) Rat anti gfap, by Thermo Fisher Scientific (1 mentions) Anti mouse f4 80, by Bio-Rad (1 mentions) Rabbit anti p akt ser473, by Cell Signaling Technology (1 mentions) Rabbit anti bdnf, by Santa Cruz Biotechnology (1 mentions) Alexa fluor 488 or, by Thermo Fisher Scientific (1 mentions) Biotinylated secondary antibody, by Jackson ImmunoResearch (1 mentions) Mouse anti myc, by Merck Group (1 mentions) Protein g sepharose beads, by Thermo Fisher Scientific (1 mentions) Irdye infrared secondary antibody, by LI COR (1 mentions) Rabbit anti phospho trkb, by Cell Signaling Technology (1 mentions) Rabbit anti myc, by Cell Signaling Technology (1 mentions) Odyssey infrared imaging system, by LI COR (1 mentions) Forskolin, by Bio-Techne (1 mentions) Rolipram, by Abcam (1 mentions) Runcaciguat, by Bayer (1 mentions) Streptavidin coated dynabeads, by Thermo Fisher Scientific (1 mentions)

5 protocols using rabbit anti trkb

Cellular Expression of TrkB and Signaling

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1) To assess the cellular source of TrkB, double immunofluorescence labeling was performed by simultaneous incubation of sections with rabbit anti-TrkB (1:100; Cell Signaling Danvers, MA, USA) overnight at 4 °C with mouse anti-NeuN (a neuronal marker; 1:100; Millipore, Billerica, MA, USA), rat anti-GFAP (an astrocyte marker; 1:200; Invitrogen, Camarillo, CA, USA), and mouse anti-F4/80 (a microglia/macrophage marker; 1:100; Serotec, Düsseldorf, Germany).
2) To assess protein expression of pTrkB, pAkt Ser473 or BDNF, double immunofluorescence labeling was performed by simultaneous incubation of sections with rabbit anti-pTrkB (1:100; Cell Signaling), rabbit anti-pAkt Ser473 (1:100; Cell Signaling) or rabbit anti-BDNF (1:100; Santa Cruz, CA, USA) overnight at 4 °C with mouse anti-NeuN (1:100; Millipore) or rat anti-GFAP (1:200; Invitrogen).
Sections were washed, followed by incubation with Alexa Fluor 488- or Alexa Fluor 594-conjugated secondary antibodies (1:500; Molecular Probes, Eugene, OR, USA) for 2 h.

Wu C.H., Chen C.C., Hung T.H., Chuang Y.C., Chao M., Shyue S.K, & Chen S.F. (2019). Activation of TrkB/Akt signaling by a TrkB receptor agonist improves long-term histological and functional outcomes in experimental intracerebral hemorrhage. Journal of Biomedical Science, 26, 53.

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Quantifying Neurotrophic Factor Signaling

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Western blots of the media and cell lysates from cultured neurons were performed using the Odyssey Infrared Imaging System (LI-COR Biosciences). Cell lysates were collected using a lysis buffer containing protease inhibitors for mammalian cell/tissue extracts (Sigma Aldrich). Media samples were immunoprecipitated with Myc antibodies bound to protein G Sepharose beads (Thermo Scientific). The following primary antibodies were used: mouse anti-Myc (Sigma Aldrich), 1:500 dilution; rabbit anti-Myc (Cell Signaling Technology), 1:1,000 dilution; mouse anti-Flag (Sigma Aldrich), 1:1,000 dilution; mouse anti-alpha tubulin (Sigma Aldrich), 1:1,000 dilution; chicken anti-TrkB (Abcam) 1:15,000 dilution; rabbit anti-TrkB (Cell Signaling Technology) 1:1,000 dilution; rabbit anti-phospho TrkB (Cell Signaling Technology) 1:1,000 dilution. Appropriate IRDYE infrared secondary antibodies (LI-COR Biosciences) or biotinylated secondary antibodies (Jackson ImmunoResearch Laboratories, Inc.) were purchased and used at a dilution of 1:10,000.

Orefice L.L., Shih C.C., Xu H., Waterhouse E.G, & Xu B. (2015). Control of Spine Maturation and Pruning through ProBDNF Synthesized and Released in Dendrites. Molecular and cellular neurosciences, 71, 66-79.

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Detailed Reagents and Materials Protocol

Cited 4 times

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BAY-747 and runcaciguat were supplied by Bayer AG (Leverkusen, Germany)^{25 (link),26 (link)}. Donepezil, an acetylcholinesterase inhibitor (AChEI), was generously donated by Abbott (Weesp, The Netherlands). Methyl 2-hydroxyethyl cellulose (Tylose^® MH300) and Tween80 (polyoxyethylenesorbitan monooleate, cat# P8074) were purchased from Sigma-Aldrich Chemie bv (Steinheim, Germany). Forskolin was purchased from Tocris Bioscience (#1099, Abingdon, UK), rolipram was purchased from Abcam (#ab120029), and sulfo-NHS-SS-biotin (#A39258) and streptavidin-coated Dynabeads (#65601) were purchased from Thermo Scientific (Bleiswijk, The Netherlands).
Mouse anti-GluA1 was purchased from Merck Millipore (#MAB2263, Burlington, MA, USA). Rabbit anti-GluA1 phospho S845 was purchased from Abcam (#ab76321), and mouse anti-GAPDH was purchased from Fitzgerald Industries (#10R-G109A, Acton, MA, US) (all three antibodies successfully used previously^{13 (link),20 (link)}). Rabbit anti-TrkB (#4603S, Cell Signaling Technologies) and mouse anti-PSD95 (#56452, QED Bioscience) were a generous gift from Bayer (as used previously^{27 (link),28 (link)}).

Nelissen E., Possemis N., Van Goethem N.P., Schepers M., Mulder-Jongen D.A., Dietz L., Janssen W., Gerisch M., Hüser J., Sandner P., Vanmierlo T, & Prickaerts J. (2022). The sGC stimulator BAY-747 and activator runcaciguat can enhance memory in vivo via differential hippocampal plasticity mechanisms. Scientific Reports, 12, 3589.

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Quantitative Western Blot Analysis of Hippocampal Proteins

Cited 1 time

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We performed western blot experiments on hippocampal tissue lysates and protein levels were quantified after separation by acrylamide gel electrophoresis (gradient 12-7.5 %) and transfer to a nitrocellulose membrane, as previously described [19 (link), 72 (link)]. Membranes were incubated overnight at 4° C with the following primary antibodies: rabbit anti-phospho NMDA receptor 2A (1:250, Abcam), mouse anti- NMDA receptor 1 (1:2000, BD Bioscience), mouse anti- PSD95 (1:1000, Abcam), mouse anti-CREB (1:750, Cell Signaling) and rabbit anti-phospho CREB (1:1000, Cell Signaling), rabbit anti-CaMKII and rabbit anti-phospho CaMKII (1:1000, Cell Signaling), rabbit anti-TrkB (1:1000, Cell signaling), rabbit anti-BDNF (1:1000, Abcam) and mouse anti-alpha-tubulin (1:30000; Sigma-Aldrich, USA). Incubations with secondary antibodies were done for 2 hours at room temperature with anti-rabbit or anti-mouse immunoglobulin G (IgG) at 1:10000 dilutions (LI-COR Biosciences GmbH, Germany). Immunoreactivity was detected with LI-COR® Odyssey® system (LI-COR Biosciences, USA) by infrared fluorescence and quantified with ImageJ 1.48v software (NIH, MA, USA) by densitometry analysis of the immunoreactive bands.

Francesca E., Kristina J., María L.L., Sarah H., Jonas W., Angel C.M, & Maioli S. (2021). Long-term exposure to polypharmacy impairs cognitive functions in young adult female mice. Aging (Albany NY), 13(11), 14729-14744.

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Multimodal Immunofluorescence Staining Protocol

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After removal from the freezer, sections were washed three times for 10 min at room temperature in Tris-buffered saline with Tween-20 (TBS-T), Tris-buffered saline (TBS) and TBS-T. Sections were then incubated overnight at 4 • C in TBS-T containing the following primary antibodies: rabbit anti-TrkB (#4606Cell Signaling Technology, Beverly, MA; 1:100), mouse anti-FLAG (#F3165 Sigma-Aldrich, St. Louis, MO; 1:200), mouse anti-NeuN (#MAB377B Millipore; 1:50), mouse anti-GFAP-biotin (#ab79203 Abcam, Cambridge, United Kingdom; 1:400), mouse anti-S100␤ (#S2532 Sigma-Aldrich; 1:500), goat anti-doublecortin (#sc-8066 Santa Cruz, Santa Cruz, CA; 1:200). After washing in TBS-T/TBS/TBS-T, sections were incubated for 2 h at 4 • C with the following secondary antibodies: donkey anti-rabbit Alexa-488 (#A-21206 Invitrogen; 1:100), donkey anti-mouse Alexa-488 (#A-21202 Invitrogen; 1:100), donkey anti-mouse Alexa-594 (#A-21203 Invitrogen; 1:100), Alexa-350 streptavidine conjugate (#S-11249 Invitrogen, 1:500), donkey anti-goat Alexa-594 (#A-11058 Invitrogen; 1:100). After washing in TBS/TBS/TBS, nuclear counterstaining was performed by 30 min incubation at room temperature with Hoechst in TBS (#33342 Sigma-Aldrich; 1:500). TUNEL staining was done using the TUNEL reaction buffer kit (Roche, Basel, Switzerland) and streptavidine-Alexa-Fluor-594 (1:2.000 in TBS-T, Invitrogen).

De Vry J., Vanmierlo T., Martínez-Martínez P., Losen M., Temel Y., Boere J., Kenis G., Steckler T., Steinbusch H.W.M., Baets M, & Prickaerts J. (2016). TrkB in the hippocampus and nucleus accumbens differentially modulates depression-like behavior in mice. Behavioural brain research, 296.

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