Cell samples were RIPA lysed buffer (P0013E, Beyotime) and protein concentration of the lysates was determined using the BCA kit (P0010S, Beyotime). Protein samples were separated by SDS-PAGE, transferred to PVDF membranes, and blocked in 10% milk in Tris-buffered saline. The membranes were then incubated at 4°C overnight in a 1:500 dilution of anti-human ALB (sc-271605, Santa Cruz), and then incubated in HRP-conjugated secondary antibody with a dilution of 1:3,000 for 2 h (BA1051, BA1055, Boster). Protein bands were visualized with an ECL kit (WBKLS0050, Merck Millipore). Data were analyzed using Image J 3.0. The gray ratio of ALB proteins and β-actin represented the relative level of the target protein. CPS1, TF, CYP3A4, CYP2D6 proteins were similarly detected by western blotting using their specific antibodies with the dilutions of 1:500, 1:10,000, 1:2,000, and 1:1,000, respectively (ab64613, ab82411, ab3572 Abcam, 93867s Cell Signaling Technology).
Ba1051
Manufactured by Boster Bio
Sourced in China
BA1051 is a laboratory equipment product. It is a multipurpose device designed for general laboratory applications. The core function of BA1051 is to assist with various experimental procedures and tasks commonly performed in a laboratory setting.
Automatically generated - may contain errors
Lab products found in correlation
Ba1054, by Boster Bio (20 mentions)
Ba1055, by Boster Bio (5 mentions)
Ripa buffer, by Beyotime (4 mentions)
Ab8245, by Abcam (4 mentions)
Pvdf membrane, by Merck Group (3 mentions)
Ripa lysis buffer, by Beyotime (3 mentions)
Bm0627, by Boster Bio (3 mentions)
Bca kit, by Beyotime (2 mentions)
Protease inhibitor pmsf, by Beyotime (2 mentions)
Nitrocellulose membrane, by Cytiva (2 mentions)
Whatman, by Cytiva (2 mentions)
Gapdh, by Proteintech (2 mentions)
Ipvh00010, by Merck Group (2 mentions)
Ab109012, by Abcam (2 mentions)
Ab51520, by Abcam (2 mentions)
Sc 271605, by Santa Cruz Biotechnology (1 mentions)
Wbkls0050, by Merck Group (1 mentions)
Ab64613, by Abcam (1 mentions)
Ab82411, by Abcam (1 mentions)
Pierce ecl western blotting substrate, by Thermo Fisher Scientific (1 mentions)
26 protocols using ba1051
1
Western Blot Analysis of Hepatic Proteins
2
Western Blot Analysis of Protein Signaling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total proteins were isolated using RIPA lysis buffer with added PMSF and phosphatase inhibitors (BEYOTIME), and were quantified using a BCA protein assay kit (BEYOTIME). The proteins were separated using 12% SDS-polyacrylamide gel electrophoresis and transferred to PVDF membranes (Millipore). After being blocked with 5% fat-free milk, the membranes were incubated with primary antibodies overnight at 4 °C. The primary antibodies used were rabbit anti-Periostin (Abcam, ab14041, 1:1000), mouse anti-β-actin (BOSTER, BM0627, 1:200), rabbit anti-ILK (Abcam, ab76468, 1:1,000), rabbit anti-pAkt (CST, 4060, 1:2,000), rabbit anti-Akt (CST, 4691, 1:1,000), rabbit anti-pGSK3β (Abcam, ab68476, 1:1,000), rabbit anti-GSK3β (Proteintech Group, Inc., 22104-1-AP, 1:1,000). Then, the membranes were incubated with HRP-conjugated goat anti-mouse IgG (BOSTER, BA1051, 1:50,000) or HRP-conjugated goat anti-rabbit IgG (BOSTER, BA1054, 1:50,000) for 2 h at 37°C. The blots were visualized with Pierce™ ECL western blotting substrate (Thermo, NCI5079) and then exposed to X-ray films (Kodak, XBT-1). The relative protein levels were quantified using Image J software (NIH, Bethesda, MD, USA).
3
Protein Extraction and Western Blotting
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total protein extraction and Western blotting were performed as described in a previous study (Wu et al. 2021 (link)). Antibodies were as follows: rabbit polyclonal antibody against FGFR2 (AF0159; Affinity, China), rabbit polyclonal antibody against TGF-βR2 (AF5449; Affinity), mouse monoclonal antibody against β-actin, horseradish peroxidase (HRP)-conjugated goat anti-rabbit second antibody (BA1054; BOSTER, China), and HRP-conjugated goat anti-mouse second antibody (BA1051; BOSTER). All antibodies were diluted 1:1000. Two-tailed t-tests were used to compare the results of different groups, and data are presented as the mean ± SEM. We used Graphpad Prism 9.0 to visualize the data.
4
Western Blot Analysis of Protein Lysates
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
N2a cells or brain tissues were lysed in RIPA buffer (Beyotime, P0013) supplemented with 1 × protease inhibitor cocktail (Roche). The total cell lysates were separated on 12–14% SDS-PAGE gels and transferred to PVDF membranes (Bio-Rad). Membranes were blocked with TBST with 5% (w/v) non-fat dry milk for 4 h (h) and probed with primary antibodies which were diluted with TBST and 5% (w/v) non-fat dry milk overnight at 4 °C. After rinsing, membranes were probed with HRP-conjugated goat anti-mouse (BA1051, Boster, Wuhan, China), goat anti-rabbit secondary antibodies (BA1055, Boster), or goat anti-mouse IgG light-chain secondary antibodies (A25012, Abbkine, Wuhan, China), then developed using BeyoECL Star kit (P0018A, Beyotime). Images were captured with an Amersham Imager 600 (GE Healthcare) imaging system. ImageJ software (National Institutes of Health, Bethesda, MD, USA) was used to quantify the intensity of protein bands.
5
Western Blot Analysis of Myogenic Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The Western blot assays were performed as previously reported [46 (link)]. The antibodies and their dilutions utilized in the Western blots are listed as follows: rabbit anti-SMARCD1/3 Ab (DF10125; Affinity Biosciences, Changzhou, China; 1:1000), rabbit anti-GAPDH (AP0063; Bioworld Technology, Bloomington, MN, USA; 1:10,000), mouse anti-MyHC (B103; DHSB, USA; 0.5 µg/mL), rabbit anti-MyoD1 (P10085; Bioss, Beijing, China; 1:500), rabbit anti-MYF5 (913349; Bioss, Beijing, China; 1:500), rabbit anti-Myogenin (P15173; Bioss, Biejing, China; 1:500), GFP-tag monoclonal antibody (AP0675M; Bioworld Technology, Bloomington, MN, USA; 1:2000), and beta-actin polyclonal antibody (Ap0060; Bioworld Technology, Bloomington, MN, USA; 1:10,000). Goat Anti-rabbit IgG-HRP (BA1054; Boster, Wuhan, China; 1:10,000) and Peroxidase-goat Anti-mouse IgG (BA1051; Boster, Wuhan, China; 1:10,000) were utilized as secondary antibodies.
6
Protein Expression Analysis Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Protein concentrations were determined using a BCA Kit (Beyotime, Beyotime Biotechnology, Shanghai, China). Equal amounts of proteins were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred to polyvinylidene fluoride membranes. Antibodies against GPR41 (66811–1-Ig, Sanying, WuHan, China) PI3K (A4992, ABclonal, Wuhan, China), phosphorylated (p)-PI3K (Ab278545, Abcam, UK), Akt (A18675, ABclonal, Wuhan, China), p-Akt (AP1208, ABclonal, Wuhan, China), mTOR (A11355, ABclonal, Wuhan, China), p-mTOR (AP0115, ABclonal, Wuhan, China), RxRα (A19105, ABclonal, Wuhan, China), SREBP1 (A5754, ABclonal, Wuhan, China), PPARγ (A16958, ABclonal, Wuhan, China), St6gal1 (A5754, ABclonal, Wuhan, China), and GAPDH (AB-P-R001, Xianzhi, Hangzhou, China) were used as primary antibodies. All primary antibodies were used at a 1:1000 dilution except for those against p-PI3K (1:500 dilution) and GPR41 (1:2000 dilution). Goat anti-rabbit-HRP (BA1054, Boster Bio, Wuhan, China) and goat anti-mouse-HRP (BA1051, Boster Bio, Wuhan, China) were used as secondary antibodies. The bands were visualized using an Enhanced Chemiluminescence (ECL) Kit (Applygen, Applygen Technologies Co., Ltd., Beijing, China). The protein bands were densitometrically analyzed using the IPP software.
7
Western Blot Analysis of ADSC-Derived Samples
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
ADSC-EVs, ADSC-NVs, cells, or tissues were lysed using RIPA buffer (Beyotime Biotechnology, P0013B) containing 1% PMSF (Aladdin, P105539), and the supernatant from lysates was used for subsequent western blot analysis. Protein samples were resolved by 10% SDS-PAGE and then transferred onto a PVDF membrane (Millipore, IPVH00010). After blocking in TBST with 5% non-fat milk for 2 h, membranes were incubated at 4 °C overnight with the following primary antibodies: CD29 (Novus, NBP2–36561), TSG101 (Servicebio, GB11618), ANG1 (Proteintech, 23,302–1-AP), VEGF (Boster Bio, PB0084), PTEN (Proteintech, 22,034–1-AP), PI3K (Proteintech, 60,225–1-AP), Akt (Affinity, AF6261), and p-Akt (Affinity, AF0016). On the next day, membranes were washed with TBST and hybridized with horseradish peroxidase (HRP)-conjugated goat anti-rabbit (Boster Biological Technology, BA1054) and anti-mouse (Boster Biological Technology, BA1051) IgG antibodies for 2 h at room temperature. Finally, bands were visualized using ECL blotting detection reagents (Servicebio, G2014).
8
Western Blot Analysis of ALB and A1AT
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The cells were dissolved in RIPA cell buffer (Beijing Solarbio Science & Technology Co., Ltd., Beijing, China), and the proteins were separated via 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) gel, which was followed by electrotransferred onto nitrocellulose membranes (EMS; Millipore, Burlington, MA, USA). The membrane was saturated with PBS containing 5% skim fat milk and 0.1% Tween 20 (PBST) for 1 hour at room temperature and incubated with the following primary antibodies: rabbit anti-human ALB (1:5,000; Proteintech; 16475-1-AP) and mouse anti-human A1AT (1:100; Proteintech; 66135-1-Ig) overnight at 4 °C. Following washing in tris-buffered saline with Tween20, the membranes were incubated with goat anti-rabbit IgG- horseradish peroxidase HRP) secondary antibody (1:10,000; Boster Bio; BA1051) or goat anti-mouse IgG-HRP secondary antibody (1:10,000; Boster Bio; BA1054) for 1 hour at room temperature. Human GAPDH was used as the control for protein loading.
9
Protein Extraction and Western Blot Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cellular protein was extracted using radio immune precipitation assay (RIPA) buffer (Beyotime, China) with phenylmethane sulfonyl fluoride (PMSF) protease inhibitor (Beyotime, China). After incubation on ice for 10 min, the supernatant was collected by centrifugation at 10,000 × g for 5 min at 4◦C. The proteins were separated in 12% SDS-PAGE and transferred onto nitrocellulose membranes (Whatman, Maidstone, UK), and then probed with antibodies following standard procedures. The antibodies and their dilutions used for western blots were as follows: Anti-PA26/SESN1 antibody (aa375-424) LS-C145280 (LS-C145280/76757; LifeSpan Biosciences, USA; 1 μg/ml), purified mouse anti-p53 (554166; BD Biosciences, USA; l:1,000), myogenin antibody (orb6492; Biorbyt, UK; 1:100), B103 (DHSB, USA; 0.5 μg/ml), p21 Cip1 antibody (GTX112898; GeneTex, USA; 1:500), Anti-gamma H2A.X (phospho S139) antibody [9F3] (ab26350; Abcam, UK; 1 μg/ml), Actived-Caspase-3 p17 polyclonal antibody (BS7004; Bioworld, USA; 1: 500) Cleaved Caspase-8 (Asp391) (18C8) Rabbit mAb (9496; Cell Signaling Technology, USA; 1:1000), Anti-Caspase-9 antibody [E23] (ab32539; Abcam, UK; 1:1000), rabbit anti-GAPDH (AB-P-R 001; Hangzhou Goodhere Biotechnology Co. Ltd., China; 1:1,500), goat anti-rabbit IgG-HRP (BA1054; Boster, China; 1:5,000) and peroxidase-goat anti-mouse IgG (BA1051; Boster, China; 1:2,500).
10
Molecular Profiling of Myo1b Signaling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After H/R irritation and/or OE-Myo1b (or si-Myo1b) transfection, total proteins of H9c2 cells were detached using RIPA Lysis Buffer (Beyotime Biotechnology). BCA assay (Beyotime Biotechnology) was performed to measure protein concentration. Then, proteins in equal concentration were electrophoresed on polyacrylamide gels and transferred onto polyvinylidene fluoride (PVDF) membrane, which were incubated with anti-Myo1b antibody (ab194356, 1 : 1000, 60 min), anti-Bcl-2 antibody (ab196495, 1 : 1000, 40 min), anti-Bax antibody (ab32503, 1 : 1000, 40 min), anti-LC3B antibody (ab63817, 1 : 500, 40 min), anti-p62 antibody (ab91526, 1 : 1000, 60 min), anti-GAPDH antibody (ab8245, 1 : 10000, 40 min, Abcam Biotechnology, CA, USA), or anti-cleaved-caspase 3 antibody (#9661, 1 : 1000, 60 min, Cell Signaling Technology, MA, USA). Then, PVDF membranes were incubated with HRP goat anti-mouse IgG (BA1051, 1 : 10000) or HRP goat anti-rabbit IgG (BA1054, 1 : 10000, Boster Biological Technology, Wuhan, China) for 60 min. Results were visualized via enhanced chemiluminescence technique. Intensities of proteins were analyzed via the Image-Pro Plus 6.0 software.
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!