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Tr 100 fj

Manufactured by Biosensis
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The TR-100-FJ is a laboratory instrument designed for the measurement and analysis of protein and nucleic acid samples. The core function of this product is to provide accurate and reliable quantification of target molecules using fluorescence detection technology.

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Lab products found in correlation

Vectashield mounting medium, by Vector Laboratories (5 mentions) Lsm 510 confocal microscope, by Zeiss (1 mentions) Potassium permanganate, by Merck Group (1 mentions) Ethanol, by Merck Group (1 mentions) Sodium hydroxide (naoh), by Merck Group (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions) Stp6000, by Leica (1 mentions) Zoletil 50, by Virbac (1 mentions)

7 protocols using tr 100 fj

1

Fluoro-Jade C Staining for Neurodegeneration

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Fluoro Jade C (FJC) is a polyanionic fluorescein derivative that selectively binds to degenerating g neuronal cell, and is more specific and sensitive than Fluoro Jade B (Schmued et al., 2005 (link)). We used Fluoro-Jade C ready-to-dilute staining kit for identifying degenerating neurons (Biosensis, TR-100-FJ), with some modification. Tissue sections were de-paraffinized, rehydrated, incubated in distilled water for 2 min. They were then incubated in a solution of potassium permanganate (1:15) for 10 min, rinsed in distilled water for 2 min and incubated in FJC solution (1:25) for 30 min. The slides were then washed and mounted on coverslips with Vecta-shield mounting medium (Vector). All sections were observed and photographed under a fluorescence microscope with a blue (450–490 nm) excitation light.
Yang L.Y., Chu Y.H., Tweedie D., Yu Q.S., Pick C., Hoffer B., Greig N, & Wang J.Y. (2015). Post-trauma administration of the pifithrin-α oxygen analogue improves histological and functional outcomes after experimental traumatic brain injury. Experimental neurology, 269, 56-66.
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2

FJC Staining for Detecting Neurodegeneration

Cited 2 times
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FJC, a derivative of polyanionic fluorescein, selectively binds to degenerating neurons. Using a FJC ready-to-dilute staining kit (TR-100-FJ, Biosensis), we identified degenerating neuronal cells in cortical tissue according to the manufacturer’s protocol with some modifications (Wang et al., 2016 (link); Yang et al., 2015 (link)). Brain sections from the different treatment groups were deparaffinized, rehydrated, and incubated in distilled water for 3 min, and then were incubated in a solution of potassium permanganate (1:15) for 10 min. They were next rinsed in distilled water for 2 min, and then incubated in the FJC solution (1:25) for 15 min. After incubation, slides were washed and mounted on coverslips with Vecta-shield mounting medium (Vector Laboratories, Burlingame, CA, USA). All sections were observed and photographed using a fluorescent inverted microscope (IX70, Olympus, Japan). Numbers of FJC-positive cells were counted in three randomly selected fields per slide with SPOT image analysis software (Diagnostic Instruments). Numbers of FJC-positive cells observed on the slides from different blinded treatment groups were counted and used to generate a mean number per treatment group.
Lin C.T., Lecca D., Yang L.Y., Luo W., Scerba M.T., Tweedie D., Huang P.S., Jung Y.J., Kim D.S., Yang C.H., Hoffer B.J., Wang J.Y, & Greig N.H. (2020). 3,6’-dithiopomalidomide reduces neural loss, inflammation, behavioral deficits in brain injury and microglial activation. eLife, 9, e54726.
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3

Fluorojade C Staining of Degenerating Neurons

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FJC, a derivative of polyanionic fluorescein, selectively binds to degenerating neurons. We used an FJC ready-to-dilute staining kit (TR-100-FJ, Biosensis) to identify degenerating neuronal cells according to the manufacturer’s protocol with some modifications. Brain sections from the different treatment groups were deparaffinized, rehydrated, and incubated in distilled water for 2 min. Brain sections were incubated in a solution of potassium permanganate (1:15) for 10 min, rinsed in distilled water for 2 min, and then incubated in the FJC solution (1:25) for 30 min. After incubation, brain slides were washed and mounted on coverslips with Vecta-shield mounting medium (Vector Laboratories, Burlingame, CA, USA). All sections were observed and photographed using a fluorescent inverted microscope (IX70, Olympus, Japan). Numbers of FJC-positive cells were counted in three randomly selected fields per slide by means of SPOT image analysis software (Diagnostic Instruments). Numbers of FJC-positive cells observed on the slides from different treatment groups were counted and used to generate a mean number per treatment group.
Huang Y.N., Yang L.Y., Greig N.H., Wang Y.C., Lai C.C, & Wang J.Y. (2018). Neuroprotective effects of pifithrin-α against traumatic brain injury in the striatum through suppression of neuroinflammation, oxidative stress, autophagy, and apoptosis. Scientific Reports, 8, 2368.
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Visualizing Degenerating Neurons with FJC

Cited 1 time
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To visualize degenerating neurons, FJC staining was performed as previously described (Yang et al., 2015 (link)). Specifically, a modified FJC ready-to-dilute staining kit was used according to the manufacturer’s instructions (Biosensis, TR-100-FJ, Thebarton, South Australia). De-waxed slides were incubated in a potassium permanganate solution (1:15) for 10 min, rinsed in distilled water for 2 min, and then incubated in FJC solution (1:25) for 30 min. The slides were thereafter washed and mounted on coverslips with Vecta-shield mounting medium (Vector, Burlingame, CA, USA). To quantify FJC-positive cells, three sections from each animal were evaluated and representative images were captured using a fluorescence microscope with a blue (450–490 nm) excitation light.
Yang L.Y., Greig N.H., Huang Y.N., Hsieh T.H., Tweedie D., Yu Q.S., Hoffer B.J., Luo Y., Kao Y.C, & Wang J.Y. (2016). Post-traumatic administration of the p53 inactivator pifithrin-α oxygen analogue reduces hippocampal neuronal loss and improves cognitive deficits after experimental traumatic brain injury. Neurobiology of disease, 96, 216-226.
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5

Fluorojade C Staining for Neurodegeneration

Cited 1 time
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A fluorojade C staining kit was used in accordance with the manufacturer’s instructions (Biosensis, TR-100-FJ, Thebarton, South Australia) and our reported studies (Fouda and Abdel-Rahman, 2017 (link), Fouda et al., 2018 (link)). Briefly, 0.06% potassium permanganate solution was used to incubate the PVN sections, on the slides, for 10 min followed by rinsing by distilled water then incubating in fluorojade C solution (1:25) for 30 min. The slides were then washed and mounted on coverslips with Vecta-shield mounting medium (Vector, Burlingame, CA, USA). A Zeiss LSM 510 confocal microscope (Carl Zeiss Inc., Thornwood, New York), and a blue (450–490 nm) excitation light was used for the visualization of stained neurons and image acquisition (Yang et al. , 2015 (link)). For quantification, Zen Lite 2011 software was used to measure the fluorescence intensity and scored by a blinded investigator.
Fouda M.A., Leffler K.E, & Abdel-Rahman A.A. (2020). Estrogen-Dependent Hypersensitivity to Diabetes-Evoked Cardiac Autonomic Dysregulation: Role of Hypothalamic Neuroinflammation. Life sciences, 250, 117598.
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6

Fluorescent Staining of Degenerating Neurons

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FJC staining was performed to identify degenerating neurons as previously described (Yu et al., 2018 (link)). According to the instruction of modified FJC ready-to-dilute staining kit (TR-100-FJ, Biosensis, Thebarton, South Australia), slides were incubated in mixture of sodium hydroxide (Sigma-Aldrich, St. Louis, MO, USA) and 70% ethanol (Sigma-Aldrich, St. Louis, MO, USA) for 5 min, followed by washing in 70% ethanol for 2 min and in distilled water for 2 min. Subsequently, slides were incubated in 0.06% potassium permanganate (Sigma-Aldrich, St. Louis, MO, USA) solution for 10 min, followed by rinsing in distilled water for 2 min. Slides then were incubated in 0.0001% FJC (MilliporeSigma, Burlington, MA, USA) for 10 min, adding 4’,6-diamidino-2-phenylindole (DAPI, 10236276001, Sigma-Aldrich, St. Louis, MO, USA) and protecting from light. Slides were then rinsed (1 min x 3) in distilled water. After dried at 50–60°C for 5 min, slides were cleared by immersing in xylene for 1 min and added coverslips with DPX mountant. The slides were visualized in blinded strategy with fluorescence microscope Leica DMi8. FJC-positive neurons (cells/mm2) were calculated in three sections per slice.
Zheng W., Matei N., Pang J., Luo X., Song Z., Tang J, & Zhang J.H. (2019). Delayed recanalization at 3 days after permanent MCAO attenuates neuronal apoptosis through FGF21/FGFR1/PI3K/Caspase-3 pathway in rats. Experimental neurology, 320, 113007.
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7

Fluoro-Jade C Staining of Apoptotic Neurons

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Mice were anesthetized with Zoletil 50 (66F4, Virbac) and Rompum solution (PP1523, Bayer) and then perfused with 4% PFA; harvested brains were cryostat sectioned at 25 μm and processed for Fluoro-Jade C (FJC) staining (TR-100-FJ, Biosensis, USA) following the manufacturer's instructions. In short, brain tissues were mounted on gelatin coated slides and heated at 57 °C for 30 min. Tissue was rehydrated with serially diluted EtOH and H2O then blocked with 0.06% KMnO4 solution for 10 min. After blocking, tissues were incubated with 0.0001% FJC solution containing 1 μg/ml DAPI for 10 min. Tissues were dehydrated with Xylene. Images were acquired using a fluorescence microscope (STP6000, LEICA). FJC positive cells were analyzed by Imager J software (Version 1.53c, National Institutes of Health, USA).
Ho M.H., Yen C.H., Hsieh T.H., Kao T.J., Chiu J.Y., Chiang Y.H., Hoffer B.J., Chang W.C, & Chou S.Y. (2021). CCL5 via GPX1 activation protects hippocampal memory function after mild traumatic brain injury. Redox Biology, 46, 102067.
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