Cells were lysed in RIRA buffer at 4°C for around 30 min followed by centrifugation at 13,200 × rpm for 10 min to remove nuclei and other cell debris. Total protein concentration was detected by the Micro BCA Protein Assay Kit (Sangon Biotech, C503061) and the lysates were either used immediately or stored at −80°C. 10–12% SDS-PAGE gels were used to separate the proteins extracted before and thereby separated proteins were completely transferred to polyvinylidene difluoride (PVDF) membranes. Five percent BSA was used to block the membranes for 1 h and the indicated primary antibodies were added and incubated overnight. Membranes were probed with the chemiluminescent detection reagents, and reactive bands were visualized using UVP ChemStudio PLUS (Analytikjena) (13 (link), 14 (link)).
Micro bca protein assay kit
Manufactured by Sangon
Sourced in China
The Micro BCA Protein Assay Kit is a colorimetric detection method for determining the total protein concentration in small sample volumes. It utilizes bicinchoninic acid (BCA) for the colorimetric detection and quantification of total protein.
Automatically generated - may contain errors
Lab products found in correlation
Microplate reader, by Thermo Fisher Scientific (2 mentions)
Uvp chemstudio plus, by Analytik Jena (1 mentions)
Cytochrome c, by Abcam (1 mentions)
γh2ax, by Cell Signaling Technology (1 mentions)
Caspase 8, by Cell Signaling Technology (1 mentions)
Caspase 3, by Cell Signaling Technology (1 mentions)
Cdkn1b p27, by Cell Signaling Technology (1 mentions)
Tubulin, by Abcam (1 mentions)
Pbcl 2 s70, by Cell Signaling Technology (1 mentions)
Caspase 9, by Cell Signaling Technology (1 mentions)
Bcl xl, by Cell Signaling Technology (1 mentions)
Mcl 1, by Cell Signaling Technology (1 mentions)
Bcl2 associated x (bax), by Cell Signaling Technology (1 mentions)
Bcl 2, by Cell Signaling Technology (1 mentions)
Cell lysis buffer, by Beyotime (1 mentions)
Dulbecco s modified eagle medium dmem medium, by Thermo Fisher Scientific (1 mentions)
3 4 5 dimethyl 2 thiazolyl 2 5 diphenyl 2 h tetrazolium bromide mtt, by Yuanye Bio-Technology (1 mentions)
Sodium dodecyl sulfate (sds), by Sangon (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Triethylamine, by Sinopharm (1 mentions)
7 protocols using micro bca protein assay kit
1
Western Blot Analysis of Protein Samples
2
Yeast Cell Protein and Oxidative Stress Quantification
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Yeasts cultured to different time points were harvested via centrifugation at 4 °C, 3000 rpm for 3 min. Cell precipitate was washed twice and resuspended with ice-cold PBS. The yeast was broken with the ultrasonic cell breaker (Ningbo Shuangjia Instrument Co., Ltd., China), 200W power for 3 s, five times of repeat at 10 s interval, the sample was placed in the ice box at all time. After broken, the cell extracts were clarified by centrifugation (10 min, 8000 rpm at 4 °C) and were used to quantify the protein content by micro BCA protein assay kit (Sangon Biotech Co., Ltd., China). Furthermore, the MDA level was assessed with enzyme-linked immunoassay kit (Jiangsu yutong biotechnology Co., Ltd., China). GSH level and SOD activity were detected by assay kits purchased from Nanjing jiancheng Bioengineering Institute (Nanjing, China).
3
Extracellular Protein Quantification via Micro BCA Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The extracellular protein concentration of the samples was detected with a micro BCA protein assay kit (C503061, Sangon Biotech, Shanghai, China). The standard curve assay solution was prepared according to the kit instructions. The bacterial solution treated with thymol at final concentrations of 0, ½ MIC, and MIC was centrifuged at 5000× g for 5 min, and the supernatant was diluted to a specific multiple. A microplate reader (Thermo Fisher Scientific, Waltham, MA, USA) was used to measure the standard and test solutions at 562 nm. Extracellular protein concentration of the samples was calculated according to the standard curve.
4
Immunoblotting of Apoptosis Regulators
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cell lysates were extracted with cell lysis buffer (Beyotime Biotechnology, Nantong, China), and the protein concentration in the lysates was quantified using a Micro BCA Protein Assay Kit (Sangon Biotech, Shanghai, China). A total of 20–50 μg of each cell lysate sample was loaded for immunoblotting and detected by antibodies that recognize Tubulin, VDAC, AIF, SMAC/DIABLO, cytochrome c (Epitomics, Hangzhou, China), CUL1 (Santa Cruz Biotechnology, Santa Cruz, CA, USA), CDKN1B/P27, CDKN2A/P16, PARP, γH2AX, caspase3, caspase8, caspase9, BCL-2, pBCL-2 (S70), BCL-XL, MCL-1, BID, BAD, BAX, and BIM (Cell Signaling Inc., Danvers, MA, USA).
5
Synthesis and Characterization of Pellethane-HA Hydrogel
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
PU (pellethane 2363-80AE) was provided by Lubrizol Corporation (Wickliffe, OH, USA). Methylenediphenyl 4,4′-Diisocyanate (MDI) was bought from Aladdin Chemical Co. Ltd. (Shanghai, China). N,N-dimethylformamide (DMF), dimethyl sulfoxide (DMSO), toluene, and triethylamine were purchased from Sinopharm Chemical Reagent Co. Ltd. (Shanghai, China), and HA was obtained from Xianding Biotechnology Co. Ltd. (Shanghai, China). 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC∙HCl), N-Hydroxysuccinimide (NHS), Lysozyme (LYS), bovine fibrinogen (BFG), human serum albumin (HSA), and 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) were obtained from Yuanye Bio-Technology Co. Ltd. (Shanghai, China). Sodium dodecyl sulfate (SDS) and the Micro BCA Protein Assay Kit were obtained from Sangon Biotech (Shanghai, China) Co. Ltd. Fetal bovine serum (FBS) and Dulbecco’s modified Eagle medium (DMEM) medium were obtained from Gibco (Grand Island, NY, USA). E. coli (DH5 alpha), S. aureus (ATCC 6538), and P. aeruginosa (ATCC 2785) and lysogenic broth (LB) medium were provided by Professor Cui, East China University of Science and Technology (ECUST, Shanghai, China). PHMG was provided by Professor Guan, ECUST.
6
Quantifying Stress Hormones in Blood and Brain
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The quantification of corticosterone (CORT) and adrenocorticotropic hormone (ACTH) in haemolysis-free blood plasma was conducted using an ELISA kit (SenBeiJia Biological Technology Co., Ltd., Nanjing, China). Total protein extracted from the hippocampus and hypothalamus samples were prepared in accordance with the instructions in the ELISA kits. Protein concentrations were determined using a Micro BCA Protein Assay kit (Sangon Biotech, Shanghai, China). The concentrations of c-Fos and brain-derived neurotrophic factor (BDNF) in the hippocampus and of CRF in the hypothalamus were measured and normalized with respect to total protein.
7
Protein Quantification of S. aureus
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The treatment of S. aureus cells was established as described in Section 3.4.3 except that the precipitated cells exposed to MEO for 12 h were tested for protein quantity with a micro BCA protein assay kit (Sangon, Shanghai, China).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!