Anti spectrin

Manufactured by Merck Group

Sourced in United States

Anti-spectrin is a laboratory reagent used to detect and quantify the presence of spectrin, a cytoskeletal protein found in various cell types. It serves as a tool for researchers and clinicians to analyze and study the structure and function of cells.

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Lab products found in correlation

Close spelling variants of this product

Spectrin (5 citations) Mouse monoclonal anti-Spectrin (4 citations) Anti-spectrin MAB1622 (3 citations) Anti-spectrin and anti-actin antibodies (2 citations) Anti-spectrin antibody (2 citations)

7 protocols using anti spectrin

Western Blot Analysis of Brain Slices

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Brain slices were stimulated with DHPG and were collected (2 slices per sample) in radioimmunoprecipitation assay (RIPA) buffer (25 mM Tris HCl pH 7.6, 150 mM NaCl, 1% NP-40, 1% sodium deoxycholate, 0.1% SDS). The samples were clarified by centrifugation at 14,000× g and the protein concentration was determined using a bicinchoninic acid assay kit (BCA; Thermo Scientific, Merelbeke, Belgium). The samples were heated for 10 min at 70 °C. A total of 10 μg of protein for each sample was loaded on a 7% SDS-polyacrylamide gel and transferred to a nitrocellulose membrane. After blocking with 5% non-fat milk, the membranes were probed overnight with anti-Arc (1/1000, Cell Signaling, Leiden, The Netherlands), anti-phospho-p44/42 MPK (pERK1/2, 1/1000, Cell Signaling Technology), anti-spectrin (1/1500, Merck Millipore, Overijse, Belgium), anti-B56α (1/250, Santa Cruz Biotechnology, Dallas, TX, USA), anti-GluA1 (1/500, Merck Millipore), anti-GluA2 (1/1000, Merck Millipore), anti-GAPDH (1/1000, Cell Signaling Technology), or anti-β tubulin (1/1000, Neuromics, Edina, MN, USA). The membranes were then incubated with secondary antibodies coupled to peroxidase (Dako, Agilent Technologies, Diegem, Belgium) and peroxidase was detected with Pierce ECL Plus (Thermo Scientific) on ECL hyperfilm.

Yerna X., Schakman O., Ratbi I., Kreis A., Lepannetier S., de Clippele M., Achouri Y., Tajeddine N., Tissir F., Gualdani R, & Gailly P. (2020). Role of the TRPC1 Channel in Hippocampal Long-Term Depression and in Spatial Memory Extinction. International Journal of Molecular Sciences, 21(5), 1712.

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Immunoblotting Analysis of Lens Proteins

Cited 1 time

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Lens proteins were subject to immunoblot analysis (Odyssey Infrared Imaging System Li-Cor, Lincoln, NE) as described [24 (link)] using anti-HSP27 (SC1049, Santa Cruz Biotechnology, Dallas, TX). Lens calpain activity was measured by immunoblot analysis of αII-spectrin cleavage products using anti-spectrin (MAB1622, EMD Millipore) and anti-β-actin (#3700, Cell Signaling Technology, CST, Danvers, MA) to control for sample loading as described [24 (link)].

Bennett T.M., Zhou Y, & Shiels A. (2016). Lens transcriptome profile during cataract development in Mip-null mice. Biochemical and biophysical research communications, 478(2), 988-993.

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Comprehensive Western Blot Analysis of Apoptosis Markers

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Western blot analysis was performed as previously reported [3] . In brief, total cellular protein concentration was determined using the BCA method then separated by SDS-PAGE and transferred to nitrocellulose (Schleicher and Schuell, Keene, NH). The primary antibodies used were rabbit monoclonal anti-XIAP, rabbit monoclonal anti-Smac/DIABLO, rabbit monoclonal anti-cytochrome c, rabbit monoclonal anti-c-IAP1, rabbit monoclonal anti-activated caspase-3, rabbit monoclonal anti-PARP, rabbit monoclonal anti-AIF, rabbit monoclonal anti-pSmad1/5, (Cell signaling Technology, MA, USA), rabbit monoclonal anti-Id1 (Calbioreagents, San Mateo, CA), rabbit polyclonal anti-Smad 1/5 (Upstate Biotechnology, NY, USA), rabbit polyclonal survivin (Novus Biologicals, CO, USA), rabbit anti-actin, an a nity isolated antigen speci c antibody (Sigma, Saint Louis, MO), rabbit polyclonal anti-GAPDH (Sigma) and mouse monoclonal anti-Spectrin (EMD Millipore, CA, USA).
Blots suggesting regulation of loading controls actin or GAPDH by Ym155 and/or JL5 were then probed with the spectrin, which is also a cytoskeletal protein.

Mondal A., NeMoyer R., Langenfeld E., Glover D., Scott M.J., Lairson L., Zloza A., Augeri D.J., Gilleran J., Peng Y., Roberge J.Y., & Langenfeld J. (2020). Bone morphogenetic protein (BMP) receptor inhibitor JL5 synergizes with Ym155 to induce Apoptosis-Inducing Factor (AIF) caspase independent cell death.

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Comprehensive Western Blot Analysis of Apoptosis Markers

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Synaptosomal Protein Quantification and Immunoblotting

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Crude synaptosomal fractions were obtained as described (Mlewski et al., 2008 (link)). All samples were valued for protein concentration (DC Protein Assay Kit, Bio-Rad Laboratory, Hercules, CA, USA) and equal protein amounts (25 μg) were separated into 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis and then transferred to Nitrocellulose membranes (Bio-Rad Laboratory, Hercules, CA, USA) as previously described (Krapacher et al., 2010 (link)). Membranes were incubated overnight at 4°C with their primary antibodies diluted in 1% skim milk in TBST. The antibodies used were anti-α-tubulin (1:3000, DM1A; Sigma-Aldrich, St. Louis, MO, USA) and anti-spectrin (1:1000, Millipore Corporation, Billerica, MA, USA). Membranes were then washed three times in TBST and incubated with a Licor secondary antibody (1:15,000, IRDye^® 800CW LI-COR Biosciences, Lincoln, NE, USA) or a horseradish peroxidase–conjugated antibody (1:2000; Jackson ImmunoResearch Laboratories, West Grove, PA, USA) for 1 h at room temperature. After two washes with TBST and two washes with TBS, bands were visualized using Odissey scanner (LI-COR Biosciences, Lincoln, NE, USA) or a chemiluminescence detection kit (ECL; Amersham Life Science, Buckinghamshire, England) according to the primary antibody used.

Ferreras S., Fernández G., Danelon V., Pisano M.V., Masseroni L., Chapleau C.A., Krapacher F.A., Mlewski E.C., Mascó D.H., Arias C., Pozzo-Miller L, & Paglini M.G. (2017). Cdk5 Is Essential for Amphetamine to Increase Dendritic Spine Density in Hippocampal Pyramidal Neurons. Frontiers in Cellular Neuroscience, 11, 372.

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Quantifying Signaling Pathway Activation

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All primary antibody solutions were prepared in TBST containing 5% bovine serum albumin at a 1:1000 dilution, except otherwise stated. The following primary antibodies were used: anti-phospho-mTOR (Ser2448) and anti-mTOR (both from Cell Signaling), anti-phospho-ERK1/2 (Thr202/Tyr204) and anti-ERK1/2 (both at 1:2000; Cell Signaling), anti-phospho-Akt (Ser473 and Thr308) and anti-Akt (all from Cell Signaling), anti-phospho-p70S6K (Thr389 from Millipore and Thr421/Ser424 from Cell Signaling), anti-p70S6K (Cell Signaling), anti-phospho-cofilin (Ser3, Abcam) and anti-cofilin (Cell Signaling), anti-phospho-PAK1/2/3 (Invitrogen) and anti-PAK3 (1:500, Millipore), anti-spectrin (1:2000; Millipore), anti-Arc (1:5000; Millipore), anti-actin (1:10000; Millipore), and anti-PTEN (Cell Signaling).

Briz V, & Baudry M. (2014). Estrogen Regulates Protein Synthesis and Actin Polymerization in Hippocampal Neurons through Different Molecular Mechanisms. Frontiers in Endocrinology, 5, 22.

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Western Blotting Analysis of Protein Markers

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Western blotting analysis was carried out according to protocols as described previously.³⁸ In brief, the total brain protein extracts from the cerebral cortex of mice with lysis buffer were prepared for Western blotting. The equivalent amount of protein was separated by 10% acrylamide denaturing gels (SDS‐PAGE) and then transferred to PVDF membrane (Millipore). Membranes were blocked with fat‐free milk for 1 hour and incubated with primary antibodies as following: anti‐β‐Actin (1:5000, Sigma‐Aldrich); anti‐Calcineurin (made by oneself); anti‐spectrin (1:1000, Millipore); anti‐SENP1 (1:2000, Abcam); anti‐FADD (1:500, Santa Cruz); anti‐Fas‐L (1:500, Santa Cruz); anti‐bcl‐2 (1:500, Santa Cruz); anti‐ZO‐1 (1:1000, Invitrogen); and anti‐Occludin (1:1000, Invitrogen) at 4ºC overnight and then incubated with HRP‐conjugated secondary antibodies (1:5000, Life Science). The proteins were visualized by an enhanced chemiluminescence detection system (Amersham Life Science). The density of the bands was quantified with ImageJ software (NIH) and normalized to β‐Actin.

Sun M., Chen X., Yin Y., Gao Y., Zhang L., Chen B., Ji Y., Fukunaga K., Han F, & Lu Y. (2020). Role of pericyte‐derived SENP1 in neuronal injury after brain ischemia. CNS Neuroscience & Therapeutics, 26(8), 815-828.

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