Samples collected from the experimental and control groups were electrophoresed using 15% polyacrylamide containing 1% sodium dodecyl sulfate (SDS) according to the Laemmli method [26 (link)]. In other words, each sample was dissolved in 1 mL of 0.01 mol/l Tris-HCL buffer solution (pH 6.8) containing 1% SDS and 25% glycerin to be used as samples for electrophoresis. After electrophoresis at 40 mA for 80 minutes under a low-voltage condition using the electrophoresis tank (Pagel AE-600, SPS-15S, ATTO Co., Japan), staining was performed with Coomassie brilliant blue (CBB, Bio-Rad, USA), followed by decoloration with stain/destain solution (Bio-Rad), and bands were detected.
Coomassie brilliant blue cbb
Manufactured by Bio-Rad
Sourced in United States
Coomassie brilliant blue (CBB) is a protein stain used in various biochemical and analytical techniques. It is a dye that binds to proteins, allowing for the visualization and quantification of protein samples. CBB is a versatile tool for protein detection and analysis in various applications such as gel electrophoresis, Western blotting, and protein assays.
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Lab products found in correlation
Polyacrylamide gel, by Nacalai Tesque (1 mentions)
Oriole fluorescent gel stain, by Bio-Rad (1 mentions)
Sds sample buffer, by Nacalai Tesque (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Amphotericin b, by Thermo Fisher Scientific (1 mentions)
Pcax app 695, by Addgene (1 mentions)
Pcax app swe ind, by Addgene (1 mentions)
4 protocols using coomassie brilliant blue cbb
1
SDS-PAGE Analysis of Protein Samples
2
Cell Proliferation Potential Assay
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To determine the proliferative potential of cells, we plated appropriate numbers of cells (1.5–5 × 103 cells) on 35‐mm dishes and grew them for 1–2 weeks. The colonies were visualized by staining with Coomassie Brilliant Blue (CBB; Bio‐Rad, Hercules, CA, USA).
3
SDS-PAGE Protein Analysis
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Elution fractions and cell lysates were boiled in sodium dodecyl sulfate (SDS) sample buffer (Nacalai Tesque, Inc.). The samples were electrophoresed on 5%–20% polyacrylamide gels under reducing condition (Nacalai Tesque, Inc.). The gels were stained using Coomassie Brilliant Blue (CBB, Bio-Rad Laboratories, Inc., Berkeley, CA) for 30 min. The gels were also stained using Oriole Fluorescent Gel Stain (Bio-Rad Laboratories, Inc.) for 2 h and were visualized with a Sayaca-Imager (DRC Co., Ltd., Tokyo, Japan).
4
Cellular Model of Alzheimer's Disease
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A cellular AD model was established in mouse neuroblastomaneuro-2a cells (ATCC-CCL-131) previously [20 (link), 21 (link)]. Among the major isoforms of APP, APP-695 is most abundant in the brain. Neuro-2a cells were transfected with pCAX vector, pCAX-APP-695 (human wild-type APP-695 isoform) or p-CAX-APP-Swe/Ind (human APP-695 K595N/M596L/V642F) (Addgene, Watertown, MA, USA). The cells were cultured and maintained in minimum essential medium (Eagle) with 2 mM L-glutamine, 0.1 mM non-essential amino acids, 2.2 g/L sodium bicarbonate, and Earle’s salt, and supplemented with 10% heat-inactivated fetal bovine serum, 100 units/mL penicillin, 100 µg/mL streptomycin, and 2.5 µg/mL amphotericin B (Thermo Fisher Scientific Inc, Waltham, MA USA). Cell culture plates were incubated at 37 °C with 5% CO2. Immunofluorescent microscopy was used to observe cell differentiation and neurite outgrowth. To determine the AD pathology in the cells, sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) was performed. One set of the gels was used for immunoblotting to check the levels of APP full-length (APP-FL), APP C-terminal fragments (APP-CTFs), and acrolein adducts; the other set was stained with Coomassie brilliant blue (CBB) (Bio-Rad Laboratories, Hercules, CA, USA) as a loading control.
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