CRC cells were washed and lysed in RIPA lysis buffer (Nanjing KeyGen Biotech Co., Ltd.). Subsequently, the protein was determined using a BCA protein content detection kit (Nanjing KeyGen Biotech Co., Ltd.) and boiled for 5 min. Denatured protein (10 µg/lane) was separated via SDS-PAGE on 10% gel, and then transferred to PVDF membranes. The membranes were blocked with TBS with Tween-20 (0.1%) containing 5% skimmed milk at room temperature for 1 h and subsequently incubated with primary antibodies (rabbit polyclonal SPARC antibody, 1:1,000, cat. no. 15274-1-AP, ProteinTech Group, Inc.; and rabbit polyclonal GAPDH antibody, 1:5,000, cat. no. 10494-1-AP, ProteinTech Group, Inc.) overnight at 4°C. The membranes were then incubated with anti-IgG secondary antibodies conjugated to horseradish peroxidase at room temperature for 1 h [HRP-conjugated Goat anti-rabbit IgG (H+L); 1:5,000; cat. no. SA00001-2; ProteinTech Group, Inc.]. Bands were visualized using the ECL reagent (Nanjing KeyGen Biotech Co., Ltd.).
Bca protein content detection kit
Manufactured by Keygen Biotech
Sourced in China
The BCA protein content detection kit is a laboratory tool used to determine the total protein concentration in a sample. It is based on the bicinchoninic acid (BCA) method, which involves the reduction of copper ions by proteins and the subsequent colorimetric detection of the resulting purple-colored complex.
Automatically generated - may contain errors
Lab products found in correlation
Revertaid first strand cdna synthesis kit, by Thermo Fisher Scientific (2 mentions)
Gapdh antibody, by Proteintech (1 mentions)
Ripa lysis buffer, by Keygen Biotech (1 mentions)
Ecl reagent, by Keygen Biotech (1 mentions)
Glut4, by Proteintech (1 mentions)
Lamin b1, by Proteintech (1 mentions)
Pgc 1α, by Proteintech (1 mentions)
Pparα, by Proteintech (1 mentions)
Pgc 1α, by Sangon (1 mentions)
Sirt1, by Sangon (1 mentions)
Power sybr green pcr master mix, by Thermo Fisher Scientific (1 mentions)
Protein extraction kit, by Beyotime (1 mentions)
Sa00001 1, by Proteintech (1 mentions)
Sa00001 2, by Proteintech (1 mentions)
Gapdh, by Proteintech (1 mentions)
β actin, by Sangon (1 mentions)
Pparα, by Sangon (1 mentions)
Gsdmd n, by Abcam (1 mentions)
Il 1β, by Boster Bio (1 mentions)
Nlrp3, by Adipogen (1 mentions)
3 protocols using bca protein content detection kit
1
Quantification of SPARC Protein Expression
2
Comprehensive Metabolic Protein Analysis
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The BCA protein content detection kit was bought from KeyGen Biotech (Nanjing, China). Hypersensitive ECL chemiluminescence kit, protease, and phosphatase inhibitor were bought from NCM Biotech (Suzhou, China). The protein extraction kit was obtained from Beyotime Institute of Biotechnology (Shanghai, China). Non-esterified fatty acids (NEFA) and blood lipid test kits were purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). Revert Aid First Strand cDNA Synthesis Kit was purchased from Thermo Fisher Scientific (Waltham, MA, United States). Power SYBR Green PCR master mix was from Invitrogen (Carlsbad, CA, United States). The primers of AMPKα, SIRT1, PGC-1α, PPARα, GLUT4, UCP3, and β-actin were designed and synthesized by Sangon Biotech (Shanghai, China). Both primary antibodies to AMPKα, PGC-1α, PPARα, GLUT4, UCP3, GAPDH, Lamin B1, and Na, K-ATPase (Cat no: 66536-1-Ig, 66369-1-Ig, 15540-1-AP, 66846-1-Ig, 10750-1-AP, 60004-1-Ig, 66095-1-Ig, and 14418-1-AP) and secondary antibodies (Cat no: SA00001-1 and SA00001-2) were purchased from the Proteintech Group (Chicago, IL, United States). The antibody to Phospho-AMPKα (Thr172, Cat #: 50081S) was purchased from the Cell Signaling Technology (Boston, MA, United States). The antibody to SIRT1 (Cat #: ab189494) was purchased from Abcam (Cambridge, United Kingdom).
3
Punicalin Attenuates NLRP3 Inflammasome
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Cell culture was purchased from GIBCO (USA); The antibodies were as follows: NLRP3 (Cat#AG-20B-0014-C100), and ASC (AG-25B-0006A) were from Adipogen, San Diego, CA, USA, Caspase-1 (Cat. No.: 22915-1-AP) was from Proteintech, Chicago, USA, GSDMD-N (ab209845) was acquired from Abcam (Cambridge, USA); The Cell Whole Protein Extraction Kit and the BCA Protein Content Detection Kit were bought from KeyGEN BioTECH (Beijing, China). Calcein-AM and PI were purchased from Tongren Chemical (Beijing, China); IL-1β and IL-18 kits were purchased from Boster Biological Technology (Wuhan, China). Punicalin (purity 98.4%, batch, 20180706) was from Chengdu Herbpurify Co. Ltd (Chengdu, China); LPS, ATP, and N-Acetylcysteine (NAC) were bought from Netotime (Beijing, China). LPS and NAC were prepared as a 0.5 μg/mL stock solution in PBS (HyClone, USA), filter-sterilized, and stored at −20°C for up to two weeks.
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