Goat-Alexa Fluor® 488 and 594 conjugated anti-mouse or anti-rabbit secondary antibodies were used (Molecular Probes, Eugene, USA). Primary antibodies included rat-anti-CD68 (Serotec, Edinburgh, UK), rabbit-anti-actin, mouse-anti-GFAP, mouse-anti-MAP-2 (Sigma Aldrich, Steinheim, Germany), mouse-anti-huntingtin (Millipore, Schwalbach, Germany), rabbit-anti-LC3 and rabbit-anti-p62 (Enzo Life Sciences, Lörrach, Germany), rat-anti-LAMP-2 (Abl93) and rat-anti-LAMP-1 (1D4B) (DSHB, Iowa City, US), rabbit-anti-LAMP-2A (Pineda, Berlin, Germany), rabbit-anti-cathepsin D (a kind gift from Prof. J. Aerts), mouse-anti-MEF2D (BD Biosciences (Heidelberg, Germany), rabbit-anti-GAPDH and rabbit-anti-α-synuclein (C-20) (Santa Cruz, Dallas, US), rabbit-anti-caspase-3, rabbit-anti-phospho-PRAS40 and rabbit-anti-PRAS40 (Cell Signalling, Frankfurt am Main, Germany) and rabbit-anti-NSE (Abcam, Cambridge, UK).
Rabbit anti nse
Manufactured by Abcam
Sourced in Germany, United Kingdom, United States
Rabbit anti-NSE is a primary antibody that recognizes neuron-specific enolase (NSE), a glycolytic enzyme expressed in neurons and neuroendocrine cells. This antibody can be used to detect and quantify NSE in various research applications.
Automatically generated - may contain errors
Lab products found in correlation
Mouse anti gfap, by Abcam (2 mentions)
Mouse anti gfap, by Merck Group (1 mentions)
Rat anti cd68, by Bio-Rad (1 mentions)
Mouse anti map2, by Merck Group (1 mentions)
Mouse anti mef2d, by BD (1 mentions)
Rabbit anti caspase 3, by Cell Signaling Technology (1 mentions)
Rabbit anti actin, by Merck Group (1 mentions)
Amersham ecl prime western blotting detection kit, by GE Healthcare (1 mentions)
Rabbit anti gapdh, by AbFrontier (1 mentions)
Multi gauge version 2, by Fujifilm (1 mentions)
Rabbit anti sucla2, by Abcam (1 mentions)
Mouse anti nestin, by Abcam (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)
Alexa fluor 594 conjugated donkey anti goat igg, by Jackson ImmunoResearch (1 mentions)
Laser confocal microscope, by Nikon (1 mentions)
Cm3050s cryostat, by Leica camera (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Yeasen (1 mentions)
Tunel kit, by Keygen Biotech (1 mentions)
4 protocols using rabbit anti nse
1
Immunohistochemistry Antibody Protocol
2
Western Blot Analysis of Protein Markers
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The isolated hippocampus and cortex samples were homogenized in buffer containing 20 mM Tris-HCl, pH 7.0, 6 M urea, 150 mM NaCl, 2 mM EDTA, and 1% Triton X-100. The homogenates were subjected to 8% SDS-polyacrylamide gel electrophoresis and analyzed by Western blot using rabbit anti-TCTP (diluted 1 : 1000, Abcam, Cambridge, MA), rabbit anti-SUCLA2 (diluted 1 : 1000, Abcam), rabbit anti-NSE (diluted 1 : 1000, Abcam), and rabbit anti-GAPDH (diluted 1 : 10,000, AbFrontier, Seoul, Korea) antibodies at 4°C overnight. The membranes were incubated with the indicated secondary antibody (diluted 1 : 5000, GE Healthcare, Madison, WI). All values were corrected with reference to the value for GAPDH, used as an internal standard. Immunoreactivity was detected by using an Amersham ECL Prime Western blotting detection kit (GE Healthcare). Western blot images were quantified using the Multi Gauge version 2.2 software (Fuji Photofilm, Tokyo, Japan).
3
Immunofluorescence Characterization of Neural Stem Cells
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Primary neural stem cells were detected and confirmed via immunofluorescence. Cultured cells were washed three times with PBS (pH 7.4) and fixed in 4% paraformaldehyde for 30 min. Cells were then permeabilized by incubation in 0.1% Triton X-100 in PBS solution. Cells were blocked with 5% goat serum for 30 min, then incubated overnight at 4 °C with primary antibodies. Antibodies used included mouse anti-Nestin (1: 500, Abcam Co., Ltd., Cambridge, UK), rabbit anti-NSE (1: 200, Abcam Co., Ltd., Cambridge, UK) and mouse anti-GFAP (1: 1000, Abcam Co., Ltd., Cambridge, UK). In the following day, cells were incubated with appropriate species-specific fluoro-conjugated secondary antibodies (Alexa Fluor 488-labeled goat anti-rabbit, goat anti-mouse and 594-labeled goat anti-rabbit IgG (H + L), 1: 1000, Sigma-Aldrich, St. Louis, MO, USA) for 2 h. Meanwhile, the nucleus was stained with DAPI (Sigma-Aldrich, St. Louis, USA). Stained cells were mounted and examined by IX 73 fluorescence microscopy (OLYMPUS). Images were acquired using cellSens Dimension software.
4
TUNEL and Immunofluorescence Staining of Rat Brains in HIBD
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TUNEL staining and double immunofluorescence staining for Bcl-2 and either NSE or GFAP were performed on the brains of rats subjected to HIBD following treatment with GFP MSCs or siIL-6 MSCs. Briefly, all rats were anaesthetised and transcardially perfused with 4% paraformaldehyde (Kelong Chemical Co., Ltd., China). Slices of brain tissue with a thickness of 12 μm were sectioned using a Leica CM3050 S cryostat. DNA fragmentation of apoptotic cells was detected using a TUNEL kit (KeyGen Biotech, China) according to the manufacturer’s instructions. The TUNEL-positive cells were counted from four randomly selected fields in each group, and the cell counts were expressed as a percentage of the total number of cells.
The sections used for double immunofluorescence staining were incubated in goat anti-Bcl-2 (1:100, Abcam, USA) together with either rabbit anti-NSE (1:100, Abcam, USA) or mouse anti-GFAP (1:100, Abcam, USA) overnight at 4 °C. The primary antibodies were visualised using Alexa Fluor 488-conjugated chicken anti-rabbit (or anti-mouse) IgG (1:100, Life Technologies, USA) and Alexa Fluor 594-conjugated donkey anti-goat IgG (1:100, Jackson, USA), and 4’,6-diamidino-2-phenylindole (DAPI, 1:100, Yeasen, China) was used to stain nuclei. The images were captured using a Nikon laser confocal microscope (Nikon, Japan).
The sections used for double immunofluorescence staining were incubated in goat anti-Bcl-2 (1:100, Abcam, USA) together with either rabbit anti-NSE (1:100, Abcam, USA) or mouse anti-GFAP (1:100, Abcam, USA) overnight at 4 °C. The primary antibodies were visualised using Alexa Fluor 488-conjugated chicken anti-rabbit (or anti-mouse) IgG (1:100, Life Technologies, USA) and Alexa Fluor 594-conjugated donkey anti-goat IgG (1:100, Jackson, USA), and 4’,6-diamidino-2-phenylindole (DAPI, 1:100, Yeasen, China) was used to stain nuclei. The images were captured using a Nikon laser confocal microscope (Nikon, Japan).
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