The sample (2 mg/mL) of WPI and MWPI were diluted 5 times with the loading buffer, centrifuged at 6000 rpm for 5 min, and then boiled for 5 min to complete denaturation. According to the protocol of the SDS-PAGE kit (Beyotime Biotechnology, Shanghai, China), the 12% separating gel and 5% concentrated gel were used for separation. Operation proceeded at 60 V for the first 30 min, and then changed to 120 V. Then, the gel was stained with 0.25% Coomassie Brilliant Blue R-250 for 30 min, and de-stained. An image was obtained using a Bio-Rad scanner (Hercules, CA, USA).
Sds page kit
Manufactured by Beyotime
Sourced in China
The SDS-PAGE kit is a laboratory equipment used for the separation and analysis of proteins based on their molecular weight. It utilizes the principle of sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) to separate protein samples in an electric field.
Automatically generated - may contain errors
Lab products found in correlation
Pvdf membrane, by Merck Group (2 mentions)
Ecl kit, by Beyotime (2 mentions)
Agarose gel, by Biowest (2 mentions)
Trizol reagent, by Tiangen Biotech (2 mentions)
Bca kit, by Beyotime (2 mentions)
Methyl thiazolyl tetrazolium (mtt), by Merck Group (2 mentions)
Sybr premix dimer eraser and reverse transcription kit, by Takara Bio (2 mentions)
Anti p tau231 ab151559, by Abcam (2 mentions)
Dna ladder, by Tiangen Biotech (2 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (1 mentions)
Bca protein quantitative kit, by Beyotime (1 mentions)
Ripa buffer, by Beyotime (1 mentions)
Pez m02 vector, by GeneCopoeia (1 mentions)
Dapi staining kit, by Beyotime (1 mentions)
Amersham imager 600, by GE Healthcare (1 mentions)
Immuno blot pvdf membrane, by Bio-Rad (1 mentions)
Pierce bca protein assay kit, by Thermo Fisher Scientific (1 mentions)
Western quick block kit, by Beyotime (1 mentions)
Amersham imager, by Cytiva (1 mentions)
Fluo 4 am, by Thermo Fisher Scientific (1 mentions)
8 protocols using sds page kit
1
SDS-PAGE Analysis of Whey Protein
2
Investigating IL-37 Mediated Modulation of Liver Cancer
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Human liver cancer cell lines, HepG2 and MHCC97H were purchased from the Cell Bank of Typical Culture Preservation Committee of Chinese Academy of Sciences (Shanghai, China), where the cell lines were authenticated via STR profiling. IL-37 gene (NM_014439) was cloned into a pEZ-M02 vector (Genecopoeia, Inc.) by the manufacturer. Lipofectamine 3000 was purchased from Invitrogen; Thermo Fisher Scientific, Inc. Rabbit anti-human IL-37 (cat. no. ab153889), NF-κB (cat. no. D14E12) and GAPDH primary antibody (cat. no. AB-P-R 001) were obtained from Abcam, Cell Signaling Technology, Inc., and Goodhere Technology, respectively. HRP-labeled goat anti-mouse IgG (cat. no. A0216), HRP-labeled Goat Anti-Rabbit IgG (cat. no. A0208), ECL kit, BCA Protein Quantitative kit, RIPA buffer, SDS-PAGE kit and DAPI staining kit were obtained from Beyotime Biotechnology. PVDF membranes were purchased from EMD Millipore.
3
Western Blot Analysis of MDSC Signaling
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Purified MDSCs from spleen or bone marrow that received various treatments were lysed on ice using RIPA lysis buffer. Cell lysates were centrifuged at 12,000g, 4 o C for 10 minutes, and the supernatant was collected to determine the protein concentration using the Pierce TM BCA protein Assay kit (Thermo Fisher Scientific). 1x sample SDS buffer was added to the supernatant for electrophoresis. 30-50µg of protein per lane alongside a prestained molecular weight protein marker was separated on an 8% SDS PAGE gel prepared from SDS-PAGE kit (Beyotime, China) and electrotransferred to immunoblot PVDF membrane (BIO-RAD, CA, USA) for protein blotting. After blocking of the membrane in western quick block kit (Beyotime, China) or 5% non-fat dry milk for at least 1 hour, it was incubated in primary antibodies against STAT3, p-STAT3, FATP2, and FATP4 with gentle agitation overnight at 4 o C. Actin was used as the housekeeping protein. HRP-conjugated secondary antibodies were used to incubate the membrane for an hour followed by protein detection with enhanced chemiluminescence (ECL) western blotting substrate and viewed on Amersham Imager 600 (GE Healthcare). Uncropped immunoblot images are shown in Figure S8.
4
SDS-PAGE Protein Analysis Reagents
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Sodium dodecyl sulfate (SDS), bovine serum albumin (BSA) and phenylmethane sulfonyl fluoride (PMSF) were purchased from Amresco (Solon, OH, USA). Phosphate-buffered saline (PBS), RIPA Lysis Buffer, SDS-PAGE kit and SDS loading buffer were obtained from Beyotime (Shanghai, China). Oligomycin A was purchased from Selleckchem (Houston, TX, USA). SNX482 was obtained from Alomone Labs (Jerusalem, Israel). Agarose, ethylenediaminetetraacetic acid (EDTA), methanol, paraformaldehyde, Tween 20, thenoyltrifluoroacetone (TTFA), polyformaldehyde, phosphatase inhibitor cocktail 3, dimethyl sulfoxide (DMSO), 4′,6-diamidino-2-phenylindole (DAPI) and N-formyl-Met-Leu-Phe (fMLP) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Fluo-4/AM, Ca2+-free HBSS, Ca2+-containing HBSS, digitonin and Coomassie blue G-250 were obtained from Invitrogen (Carlsbad, CA, USA). Unless otherwise stated, all concentrations shown are the final concentrations.
5
Oxidative Stress and Autophagy in Human Bronchial Epithelial Cells
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Human bronchial epithelial cells and lentiviruses were purchased from GeneChem. FBS, RPMI‐1640 medium and penicillin/streptomycin were obtained from HyClone. Commercial kits used for the analysis of CAT, MDA, SOD and GSH‐Px were purchased from Nanjing Jiancheng Bioengineering Institute. The 2′7′‐dichlorofluorescein diacetate (DCFH‐DA) used to measure reactive oxygen species (ROS) formation was obtained from Beyotime Biotechnology. A cell proliferation assay kit (MTS method) was obtained from Promega Biotech Company. TRIzol used for RNA extraction, a PrimeScript RT reagent kit, a SYBR Premix TaqTM kit and dT primers were all purchased from TaKaRa Biotechnology Company. Total protein extraction kits were purchased from Invent Biotechnologies Institution. A BCA reagent kit and SDS‐PAGE kit were obtained from Beyotime Biotechnology. PVDF membranes were purchased from Millipore. Antibodies against IP3R and fluorescently labelled secondary antibody were obtained from Cell Signaling Technology. Antibodies against Atg5, LC3, P62, Beclin1 and β‐actin were purchased from CST, and goat anti‐rabbit IgG HRP secondary antibody was obtained from the Absin Bioscience Institution. 3‐Methyladenine (3‐MA) was purchased from MedChemExpress.
6
Optimized Molecular Techniques for Neurological Research
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The DNA kit was purchased from Foregene Company (Chengdu, Sichuan, China). The MTT (3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide) was bought from Sigma-Aldrich (St. Louis, MO, USA). The PCR primers were synthesized by and purchased from Genscript Company (Nanjing, Jiangsu, China). The agarose gel was purchased from Biowest Company. The TRIzol ® Reagent and DNA ladder were acquired from TIANGEN (Tianjin, China). The SYBR Premix Dimer Eraser and Reverse Transcription kit were purchased from Takara (Japan). The 40 μm cell filters were obtained from Biologix Company (Shanghai, China). The Lipofectamine 3000 reagent was obtained from Invitrogen (Waltham, MA, USA). The BSA was purchased from Biofroxx Biotechnology (Germany). The BCA kit and SDS-PAGE kit were purchased from Beyotime Biotechnology (Shanghai, China). The anti-BMP4 (ab39973) and anti-P-tau231 (ab151559) antibodies were obtained from Abcam (Cambridge, MA, USA). The anti-APP (#29765), anti-Tau (#4019), and anti-P-Tau181 (#12885) antibodies were obtained from Cell Signaling Technology (Danvers, MA, USA). The anti-BAX (50599-2-Ig), anti-Bcl-2 (26593-1-AP), and anti-PSEN1 (16163-1-AP) antibodies were obtained from ProteinTech Company (Wuhan, Hubei, China).
7
HepG2 Protein Extraction and Western Blot Analysis
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HepG2 protein was extracted using the SDS-PAGE kit (Beyotime Biotechnology, Shanghai, China). Protein concentrations were determined using a BCA protein assay kit (Shanghai Fuheng Biotechnology). The protein was then separated using SDS-PAGE and transferred to a polyvinylidene fluoride membrane. After blocking it for one hour with 5% skim milk powder, the primary antibody (anti-LC3, Proteintech Group, and b-actin, Zen-Bio) was added and shaken overnight at 4 °C. Secondary antibody (Zen-Bio) was then added and cells were subjected to agitation at room temperature for one hour. The chemiluminescence assay was carried out using the ECL kit (Shanghai Biyuntian Biotechnology) and detected by ChemiDoc XRS+ (Bio-Rad). Data were analyzed with the ImageJ program.
8
Optimized Neurodegeneration Molecular Analysis
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DNA kit was purchased from Foregene Company (Chengdu, Sichuan, China). MTT (3-(4,5-dimethyl-2thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide) was bought from Sigma-Aldrich (St. Louis, MO, USA).
PCR primer synthesis was purchased from Genscript Company (Nanjing, Jiangsu, China). Agarose gel was purchased from Biowest Company (France). TRIzol ® Reagent and DNA ladder were acquired from TIANGEN (Tianjin, China). SYBR Premix Dimer Eraser and Reverse transcription kit was purchased from Takara (Japan). 40μm cell lters were obtained from Biologix Company (Shanghai, China). Lipofectamine 2000 reagent was obtained from Invitrogen (Waltham, MA, USA). BSA was purchased from Biofroxx Biotechnology (Germany). BCA kit and SDS-PAGE kit were purchased from Beyotime Biotechnology (Shanghai, China). Anti-BMP4 (ab29973) and anti-P-tau231 (ab151559) antibodies were obtained from Abcam (Cambridge, MA, USA). Anti-APP (#P05067), anti-Tau (#P10636-8), anti-P-Tau181 (#P10636-8) antibodies were obtained from Cell Signaling Technology (Danvers, MA, USA). Anti-BAX (50599-2-Ig), anti-Bcl-2 (26593-1-AP), anti-PSEN1 (16163-1-AP) and anti-Ab42 (25524-1-AP) antibodies were gained from ProteinTech Company (Wuhan, Hubei, China).
PCR primer synthesis was purchased from Genscript Company (Nanjing, Jiangsu, China). Agarose gel was purchased from Biowest Company (France). TRIzol ® Reagent and DNA ladder were acquired from TIANGEN (Tianjin, China). SYBR Premix Dimer Eraser and Reverse transcription kit was purchased from Takara (Japan). 40μm cell lters were obtained from Biologix Company (Shanghai, China). Lipofectamine 2000 reagent was obtained from Invitrogen (Waltham, MA, USA). BSA was purchased from Biofroxx Biotechnology (Germany). BCA kit and SDS-PAGE kit were purchased from Beyotime Biotechnology (Shanghai, China). Anti-BMP4 (ab29973) and anti-P-tau231 (ab151559) antibodies were obtained from Abcam (Cambridge, MA, USA). Anti-APP (#P05067), anti-Tau (#P10636-8), anti-P-Tau181 (#P10636-8) antibodies were obtained from Cell Signaling Technology (Danvers, MA, USA). Anti-BAX (50599-2-Ig), anti-Bcl-2 (26593-1-AP), anti-PSEN1 (16163-1-AP) and anti-Ab42 (25524-1-AP) antibodies were gained from ProteinTech Company (Wuhan, Hubei, China).
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