Ab5392

Manufactured by Agilent Technologies

Sourced in United States, United Kingdom

The Ab5392 is a compact and versatile laboratory instrument designed for general analytical applications. It features a high-performance detector and advanced data processing capabilities to enable accurate and reliable measurements. The core function of the Ab5392 is to provide precise analytical results for a variety of sample types.

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Lab products found in correlation

Pierce bca protein assay kit, by Thermo Fisher Scientific (2 mentions) Odyssey clx infrared imaging system, by LI COR (1 mentions) Z033429, by Agilent Technologies (1 mentions) Iblot 2 dry blotting system, by Thermo Fisher Scientific (1 mentions) Bolt 4 12 bis tris plus gel, by Thermo Fisher Scientific (1 mentions) Ab51502, by Abcam (1 mentions) Ab31940, by Abcam (1 mentions) Ab18465, by Abcam (1 mentions) Z033429 2, by Abcam (1 mentions) Dapi fluoromount g, by Southern Biotech (1 mentions)

3 protocols using ab5392

Western Blot Analysis of Neuronal and Glial Markers

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Cells were lysed in RIPA buffer with protease inhibitor, and total protein was extracted and quantified (Pierce BCA Protein Assay Kit, Thermo Scientific). Total protein (20µg) was separated using a Bolt 4–12% Bis‐Tris Plus Gel (Life Technologies) and transferred to a nitrocellulose membrane using an iBlot2 dry blotting system (Thermo Scientific). Membranes were blocked for 1–4 h (Rockland Immunochemicals, VWR International, Arlington Heights, IL, USA); primary antibodies (rabbit anti‐Synapsin1, EMD‐Millipore AB1543P, 1:1,500; mouse anti‐PSD‐95, Neuromab, 1:1,500; chicken anti‐MAP2, Abcam ab5392, 1:2,000; rabbit anti‐GFAP, DAKO Z033429, 1:2,000) in blocking buffer incubated, shaking, overnight at 4°C and secondary antibodies (IRDye 680RD and IRDye 800CW, 1:5,000 in blocking buffer) for 1 h at room temperature. Proteins were detected by an Odyssey CLx infrared imaging system (LiCOR Biosciences, Lincoln, NE, USA), and semi‐quantitative analysis of signal intensity was corrected to the relative quantification of β‐actin.

Trujillo C.A., Adams J.W., Negraes P.D., Carromeu C., Tejwani L., Acab A., Tsuda B., Thomas C.A., Sodhi N., Fichter K.M., Romero S., Zanella F., Sejnowski T.J., Ulrich H, & Muotri A.R. (2020). Pharmacological reversal of synaptic and network pathology in human MECP2‐KO neurons and cortical organoids. EMBO Molecular Medicine, 13(1), e12523.

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Immunofluorescence Staining of Neuronal Cultures

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Neuronal cultures grown on glass coverslips were fixed with a solution of 4% paraformaldehyde and 4% sucrose in 1× PBS pH 7.4 for 8 min at 27 °C, blocked for 1 h at 4 °C with 0.1% (w/v) gelatin and 0.3% (v/v) Triton X-100 in 1× PBS pH 7.4 (GDB), and incubated overnight at 4 °C with the following primary antibodies diluted in GDB: anti-GFP (rabbit, 1:500, Life Technologies, A21311), anti-MAP2 (chicken, 1:1,000, Lifespan Biosciences, LS-C61805) and anti-MAP2 (chicken, 1:1,000, Abcam AB5392), anti-GFAP (rabbit, 1:500, Dako, Z033429-2), anti-SATB2 (mouse, 1:500, Abcam AB51502), anti-CTIP2 (rat, 1:300, Abcam, AB18465), anti-TBR1 (rabbit, 1:300, Abcam, AB31940). The OSTN antibody (rabbit, 1:500) was raised against a C-terminal region of OSTN (NP_937827.1, amino acids 112–127, PKRRFGIPMDRIGRNR), then affinity-purified. All secondary antibodies were AlexaFluor-conjugated (Life Technologies). Coverslips were mounted with DAPI Fluoromount G (SouthernBiotech).

Ataman B., Boulting G.L., Harmin D.A., Yang M.G., Baker-Salisbury M., Yap E.L., Malik A.N., Mei K., Rubin A.A., Spiegel I., Durresi E., Sharma N., Hu L.S., Pletikos M., Griffith E.C., Partlow J.N., Stevens C.R., Adli M., Chahrour M., Sestan N., Walsh C.A., Berezovskii V.K., Livingstone M.S, & Greenberg M.E. (2016). Evolution of Osteocrin as an activity-regulated factor in the primate brain. Nature, 539(7628), 242-247.

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Immunocytochemistry for Neural Stem Cell Differentiation

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To assess the differentiation of neural stem cells to neurons and astrocytes the immunocytochemistry was performed with MAP-2 antibody (for detecting neurons) and GFAP (for detecting astrocytes). Briefly, the cells were fixed with paraformaldehyde 4% at 4℃ for 20 minutes, followed by washing with phosphate buffer saline (PBS). The primary antibodies for MAP-2 (Abcam, Cambridge, UK; ab5392 1:1000) and GFAP (DAKO, Glostrup, Denmark; catalogue number Z0334 1:700) in 5% goat serum and 0.01% Triton-X100 were added and the cells were kept at room temperature for 1 hour. The primary antibodies were then removed and the cells were twice washed with PBS and the secondary antibodies were added for 1 hour in room temperature. For final step, the cells were washed three times with PBS and after that visualized and photographed under a fluorescent microscope (model Nikon X66; Nikon, Tokyo, Japan).

Hosseini S.M., Sharafkhah A., Koohi-Hosseinabadi O, & Semsar-Kazerooni M. (2016). Transplantation of Neural Stem Cells Cultured in Alginate Scaffold for Spinal Cord Injury in Rats. Asian Spine Journal, 10(4), 611-618.

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