Sc 17839

Manufactured by Santa Cruz Biotechnology

Sourced in United States, Canada

Sc-17839 is a laboratory reagent product offered by Santa Cruz Biotechnology. It is a small molecule compound with a specific catalog number. The core function of this product is to serve as a research tool for scientific investigations, though its exact intended use is not provided in this unbiased, factual description.

Automatically generated - may contain errors

Lab products found in correlation

Sp5 2 confocal microscope, by Leica (1 mentions) Alexa fluor 488, by Thermo Fisher Scientific (1 mentions) Alexa 594, by Thermo Fisher Scientific (1 mentions) Sc 15081, by Santa Cruz Biotechnology (1 mentions) Aim software, by Zeiss (1 mentions) Lsm 700 confocal microscope system, by Zeiss (1 mentions) Ab180655, by Abcam (1 mentions) Ab32216, by Abcam (1 mentions) Odyssey system, by LI COR (1 mentions) Nb100 56745, by Novus Biologicals (1 mentions) Irdye 800cw goat anti mouse, by LI COR (1 mentions) Irdye 680rd goat anti rabbit, by LI COR (1 mentions) Bca assay kit, by Thermo Fisher Scientific (1 mentions) Sc 52, by Santa Cruz Biotechnology (1 mentions) Complete ultra tablet, by Roche (1 mentions) Mouse anti β actin, by Abcam (1 mentions) Ab8227, by Abcam (1 mentions) Af7021, by Affinity Biosciences (1 mentions) Bca reagent, by Thermo Fisher Scientific (1 mentions) Cofilin, by Cell Signaling Technology (1 mentions)

8 protocols using sc 17839

Immunostaining of Neuronal Culture

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For immunostaining, neurons were cultured in poly-d-lysine-coated coverslips and treated with TNI, glutamate, and/or edonerpic maleate. After being fixed with 4% paraformaldehyde, permeabilized with 0.1% Triton X-100 and washed with PBS three times, neurons were blocked by 5% bovine serum albumin (BSA). Incubation with the DMPO (1:50, ab104902, Abcam) and Arc (1:50, sc-17839, Santa Cruz) primary antibodies was performed at 4 °C overnight. After being washed by PBS with Tween-20 (PBST) three times, the samples were incubated with the secondary antibody at 37 °C for 1 h. Then, incubation with 4′, 6-diamidino-2-phenylindole (DAPI) was performed to stain the nuclei, and the images were obtained using a Leica SP5 II confocal microscope.

Chen T., Yang L.K., Ai P., Zhu J., Hang C.H, & Wang Y.H. (2022). Edonerpic maleate regulates glutamate receptors through CRMP2- and Arc-mediated mechanisms in response to brain trauma. Cell Death Discovery, 8, 95.

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Investigating Arc and 5HT2C Receptor Expression

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Animals received systemic injections of either saline or citalopram as described above. 45 minutes after fear conditioning, animals were given an overdose of sodium pentobarbital (100 mg/kg), and then perfused transcardially with 0.9% saline and 4% paraformaldehyde in 0.1M phosphate buffer (PB). Tissue was post-fixed for 4 hours then transferred to a 20% sucrose solution. Tissue was sliced into 80 µM coronal sections using a Vibratome. Every other slice was collected, so there was no need to correct for double counting. 5 or 6 slices per animal were collected, covering the rostro-caudal extent of the BNSTov. Tissue was washed in 0.1M PB 3 times, then in PB with 1% Triton (PBT) 3 times for 5 min each wash. Slices were blocked in 1% bovine serum albumin (BSA; Sigma Fraction V, A-3059) in PBT for 1 hr and incubated for 48 hours at 4 degrees C in primary antibodies in BSA. Primaries used were a mouse monoclonal anti Arc/Arg3.1 antibody (1:250; Santa Cruz Biotechnology, sc-17839) and a goat polyclonal antibody against the 5HT2C receptor (1:1000; Santa Cruz Biotechnology, sc-15081). Slices were washed in PBT and incubated in secondary antibodies in PBT for 1 hr. Secondary antibodies used were donkey anti-mouse Alexa Fluor-488, and donkey anti-goat Alexa-594 (1:200; Life Technologies). Slices were washed in PBS, mounted on slides and coverslipped.

Pelrine E., Diana Pasik S., Bayat L., Goldschmiedt D, & Bauer E.P. (2016). 5-HT2C receptors in the BNST are necessary for the enhancement of fear learning by selective serotonin reuptake inhibitors. Neurobiology of learning and memory, 136, 189-195.

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Immunofluorescence Quantification of Arc in Hippocampus

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Immunofluorescence staining was performed on the sectioned hippocampus (n = 10 per group) to assess the expression of Arc. Hippocampus sections were blocked with 5% goat serum for 30 min at room temperature, then incubated with primary anti-Arc (sc-17839, 1:100, Santa Cruz, CA, USA) and anti-NeuN (1:100, ab12763, Abcam) overnight at 4 °C. The secondary antibodies used were anti-mouse and anti-rabbit respectively (1:1000; Invitrogen, Carlsbad, CA, USA). Sections were incubated with the secondary antibodies in the dark for 2 h at room temperature. The nuclei were then stained with DAPI. The anti-NeuN was used as neuronal marker. The expression level of Arc was assessed by fluorescence microscopy. Confocal images of sections were sequentially acquired with Zeiss AIM software on a Zeiss LSM 700 confocal microscope system. The analysis method is the same as ROS mentioned above.

Li T., Xu G., Yi J, & Huang Y. (2022). Intraoperative Hypothermia Induces Vascular Dysfunction in the CA1 Region of Rat Hippocampus. Brain Sciences, 12(6), 692.

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Quantitative Protein Analysis of Calpain-1 KO Mice

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Whole brains were homogenized in RIPA buffer with protease inhibitors at 4°C. After centrifugation at 13,000 × g at 4°C for 15 min, protein amounts in the supernatant were quantified using the BCA Assay kit (Pierce Biotechnology). Proteins from whole brains of WT and calpain-1 KO mice were subjected to 10% SDS-PAGE, and proteins were transferred to a PVDF membrane with 100 V for 1 h at 4°C. After blocking for 2 h at room temperature with 3% bovine serum albumin in TBS buffer, membranes were incubated at 4°C overnight with rabbit anti-HSPA1B (1:500; PA5-28369; Thermo Fisher Scientific), anti-DNAJB1 (1:1000; 13174-1-AP; Proteintech), anti-Insulin degrading enzyme/IDE (1:1000; ab32216; abcam), anti-PLA2G4E (1:200; 18088-1-AP; Proteintech), anti-NGFI-B alpha/Nur77/NR4A1 (1:1000; NB100-56745; Novus Biologicals), anti-PER2 (1:300; ab180655; Abcam) antibodies and mouse anti-ARC (1:500; sc-17839; Santa Cruz) antibody. After incubation in primary antibodies, membranes were washed with TBST buffer and incubated for 2 h at room temperature with IRDye 680RD goat anti-rabbit (1:10,000; LI-COR Biosciences) and IRDye 800CW goat anti-mouse (1:10,000; LI-COR Biosciences). Thereafter, membranes were washed 3 times with the TBST and 1 time with TBS buffer. Immunoreactivity was detected with the LI-COR Odyssey system (LI-COR Biosciences).

Su W., Zhou Q., Wang Y., Chishti A., Li Q.Q., Dayal S., Shiehzadegan S., Cheng A., Moore C., Bi X, & Baudry M. (2020). Deletion of the Capn1 Gene Results in Alterations in Signaling Pathways Related to Alzheimer’s Disease, Protein Quality Control and Synaptic Plasticity in Mouse Brain. Frontiers in Genetics, 11, 334.

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Immunolabeling of Neuronal Markers

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Cells were fixed with a solution containing 4% paraformaldehyde (PFA), 4% sucrose, and PBS for 15 min at room temperature (RT), blocked with a solution containing 10% goat serum, 2% bovine serum albumin (BSA), and PBS for 1 h at RT. Rabbit-anti-NeuN (Thermo Fisher Scientific catalog #711054, RRID:AB_2610583), mouse-anti-Arc (C7; Santa Cruz Biotechnology catalog #sc-17839, RRID: AB_626696), and rabbit-anti-c-Fos (Santa Cruz Biotechnology catalog #sc-52, RRID:AB_2106783) primary antibody were used at 1:300. Primary antibodies were incubated for 1 h at RT in a dilution buffer containing 1:1 block solution and PBS-Triton X-100 solution at 1:300. Dishes were washed three times with PBS-Triton X-100 and incubated with Alexa Fluor 488-conjugated secondary antibody 1:1000 (Invitrogen-Invitrogen) in dilution buffer for 1 h at RT. Washing was repeated as per the above. The cells were subsequently incubated with 5 μm DAPI for 5 min before being imaged.

Jiang Y, & VanDongen A.M. (2021). Selective Increase of Correlated Activity in Arc-Positive Neurons after Chemically Induced Long-Term Potentiation in Cultured Hippocampal Neurons. eNeuro, 8(6), ENEURO.0540-20.2021.

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Hippocampal Slice Preparation and Analysis

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Hippocampal slices were cut (300 µm) with a McIlwain tissue chopper (Mickle, UK) and incubated for at least 1 h at room temperature in oxygenated (95% O₂, 5% CO₂) ACSF as described^{39 (link),40 (link)}. They were kept at 32 °C for another hour before treatment with DHPG (100 µM) for 5 min or with Sal003 (20 µM) for 10 min, before snap-freezing over dry ice. In all instances, similar slices were treated with vehicle as control. Frozen slices were lysed in homogenizing buffer (200 mM HEPES, 50 mM NaCl, 10% glycerol, 1% Triton X-100, 1 mM EDTA, 50 mM NaF, 2 mM Na₃VO₄, 25 mM β-glycerophosphate and EDTA-free complete ULTRA tablets (Roche, Indianapolis, IN)). Western blotting was performed as described by Huang et al.^40.Primary antibodies for western blotting were rabbit anti–p-eIF2α (Ser51) (1: 500, 3398, Cell Signaling and Technology Laboratories, Danvers, MA), mouse anti-total eIF2α (1: 1,000, 2103, Cell Signaling), mouse anti-β-actin (1:10,000, ab8227, Abcam, Cambridge, MA), mouse anti-OPHN1 (1:1,000, sc-374330, Santa Cruz Biotechnology, Santa Cruz, CA) and mouse anti-Arc (1: 200, sc-17839, Santa Cruz Biotechnology). Secondary antibodies for western blotting were peroxidase-conjugated goat anti-rabbit (1:10,000, 111–035–144, Jackson ImmunoResearch) and goat anti-mouse (1:10,000, 115–035–003, Jackson ImmunoResearch).

Viana Di Prisco G., Huang W., Buffington S.A., Hsu C.C., Bonnen P.E., Placzek A.N., Sidrauski C., Krnjević K., Kaufman R.J., Walter P, & Costa-Mattioli M. (2014). Translational control of mGluR-dependent long-term depression and object-place learning by eIF2α. Nature neuroscience, 17(8), 1073-1082.

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Western Blot Analysis of Neuronal Markers

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Neurons from different groups of cultures were collected and lysed with RIPA lysis buffer supplemented with protease inhibitors for 30 min at 4°C. Then, the samples were centrifuged at 13000 rpm, 4°C for 15 min. The supernatant was collected and boiled in a loading buffer. Protein concentration was measured using a BCA reagent (Thermo Scientific, USA) and then transferred to a polyvinylidene difluoride (PVDF) membrane. The membrane was then incubated overnight at 4°C with following primary antibodies: anti-Fos (1:1000, #AF0132, Affinity), anti-Arc (1:250, #sc-17839, Santa Cruz), and anti-GAPDH (1:1000, #AF7021, Affinity). Immunoglobulin G horseradish peroxidase (HRP)-labeled secondary antibodies (1:5000) were then applied at the room temperature for additional 2 h. Millipore's enhanced chemiluminescence was used to visualize the proteins. The data were analyzed using ImageJ software (National Institutes of Health, Bethesda, MD, USA).

Liu X.Y., Song R.H., Li T., Tan X., Zhang X.H., Pang K.K., Shen J.Y., Yue Q.W, & Sun J.H. (2021). Nicotinamide adenine dinucleotide promotes synaptic plasticity gene expression through regulation N-methyl-D-aspartate receptor/Ca²⁺/Erk1/2 pathway. The Chinese journal of physiology, 64(6).

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Western Blot Analysis of Neural Markers

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Tissues (optic nerve, hippocampus, cortex and corpus callosum) were micro-dissected and homogenized with RIPA lysis buffer. Equal amounts of proteins were loaded and separated by SDS-PAGE and transferred to PVDF membranes. Membranes were blocked in 0.1% Tween20 PBS buffer containing 5% BSA overnight at 4°C and incubated with primary antibodies followed by anti-mouse or anti-rabbit IgG conjugated to HRP. Enhanced chemiluminescence was performed with a West Pico Kit and detected by FluorChem E system (ProteinSimple, USA). The following primary antibodies were used: Cdk5 (sc-173, Santa Cruz, 1:1000); PLP (ab28486, Abcam, 1:1000); Caspr (MABN69, Millipore, 1:1000); MBP (SMI-99P, Covance, 1:1000); CREB (#9197, Cell signaling, 1:1000) and p-CREB (#9198, Cell signaling, 1:1000); Cofilin (#5175, Cell signaling, 1:1000) and p-Cofilin (#3311, Cell signaling, 1:1000); Fos (#4384, Cell signaling, 1:1000) and Arc (sc-17839, Santa Cruz, 1:1000) and β-actin (sc-47778, Santa Cruz, 1:1000). The density of immunoblots were quantified against normalized internal loading control using ImageJ software.

Luo F., Zhang J., Burke K., Romito-DiGiacomo R.R., Miller R.H, & Yang Y. (2018). Oligodendrocyte-specific loss of Cdk5 disrupts the architecture of nodes of Ranvier as well as learning and memory. Experimental neurology, 306, 92-104.

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