Stock solutions of NCTD (Yuanye Bio-Technology Co., Ltd., Shanghai, China), 5-FU (Selleck Biotech Co., Ltd., Houston, TX, USA), and DMSO (Sigma-Aldrich, St. Louis, MO, USA) were prepared. The preparation of NCTD and 5-FU stock solutions: 20 mg NCTD powder was diluted to 100 mM with 1.2 mL DMSO. The working concentration was 12.5/25 μM. Analogously, 20 mg 5-FU powder was diluted to 100 mM with 1.5 mL DMSO. The working concentration of 5/10 μM was then prepared. Dulbecco’s modified Eagle’s medium (DMEM), fetal bovine serum (FBS), penicillin/streptomycin (PS), and trypsin were obtained from Gibco, Thermo Fisher Scientific, Inc. (Waltham, MA, USA). β-actin monoclonal antibodies were purchased from Proteintech Group, Inc. (Rosemont, IL, USA). Antibodies against caspase-3 and caspase-9 were purchased from Cell Signaling Technology Inc. (Boston, MA, USA). Goat anti-rabbit IgG-HRP and goat anti-mouse IgG-HRP were purchased from Jackson ImmunoResearch Inc. (West Grove, PA, USA). Cell counting kit-8 (CCK-8) was purchased from Beyotime Biotechnology (Shanghai, China). The Annexin V-FITC/PI Apoptosis Detection Kit and TUNEL FITC Apoptosis Detection Kit were purchased from Vazyme Biotech Co., Ltd. (Nanjing, China).
β actin monoclonal antibody
Manufactured by Proteintech
Sourced in United States, China
The β-actin monoclonal antibody is a laboratory reagent used to detect the presence and quantify the levels of the β-actin protein in biological samples. β-actin is a widely expressed and highly conserved cytoskeletal protein that plays a crucial role in various cellular processes.
Automatically generated - may contain errors
Lab products found in correlation
Trypsin edta, by Thermo Fisher Scientific (3 mentions)
Trypsin, by Thermo Fisher Scientific (2 mentions)
4 6 diamidino 2 phenylindole (dapi), by Beyotime (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Antibodies against caspase 3 and caspase 9, by Cell Signaling Technology (1 mentions)
Goat anti mouse igg hrp, by Jackson ImmunoResearch (1 mentions)
Goat anti rabbit igg hrp, by Jackson ImmunoResearch (1 mentions)
Penicillin streptomycin p s, by Thermo Fisher Scientific (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Cell counting kit 8 cck 8, by Beyotime (1 mentions)
Annexin 5 fitc pi apoptosis detection kit, by Vazyme (1 mentions)
Tunel fitc apoptosis detection kit, by Vazyme (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Glutamine, by Thermo Fisher Scientific (1 mentions)
Mepacrine, by Merck Group (1 mentions)
Fitc phalloidin, by Merck Group (1 mentions)
Hsp27, by Cell Signaling Technology (1 mentions)
Total p38, by Cell Signaling Technology (1 mentions)
Phospho p38, by Cell Signaling Technology (1 mentions)
Luciferin luciferase, by Chrono-Log (1 mentions)
14 protocols using β actin monoclonal antibody
1
Anticancer Drug Stock Solution Preparation
2
Porcine Alveolar Macrophage Cell Line Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The porcine alveolar macrophage cell line 3D4/21 (ATCC CRL-2843) [17 (link)] and M. hyopneumoniae strain JS were kind gifts from the Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences, China; antibiotics (streptomycin and penicillin), fetal bovine serum, trypsin, glutamine, fetal rat serum (FBS), and roswell park memorial institute (RPMI)-1640 medium + 25 mM N-piperazine-N’-[2-ethanesufonic acid] (HEPES) were purchased from Gibco (Grand Island, NY, USA). CYP3A29 and β-Actin monoclonal antibodies were purchased from Proteintech Group, Inc. (Chicago, IL, USA). Monoclonal antibodies against PXR were purchased from Sigma-Aldrich (St. Louis, MO, USA). All the other chemicals were of analytical grade.
3
Platelet Activation Signaling Pathways
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Collagen, thrombin, ADP and luciferin/luciferase were provided by the Chrono-Log Corporation. FITC-phalloidin, H2O2 and mepacrine were purchased from Sigma-Aldrich; Merck KGaA. Antibodies against phospho-ERK1/2 (catalog no. 4370S), phospho-p38 (catalog no. 4511S), phospho-HSP27 (catalog no. 2401S), total-p38 (catalog no. 8690S), total-ERK (catalog no. 4695S) and HSP27 (catalog no. 95357S) were purchased from Cell Signaling Technology Inc. β-actin monoclonal antibody (catalog no. 66009-1-Ig) was obtained from ProteinTech Group, Inc. The antibody for integrin β3 (D-11) (catalog no. sc-365679) was obtained from Santa Cruz Biotechnology Inc. PAC-1 antibodies (catalog no. MA5-28564) were from Invitrogen; Thermo Fisher Scientific, Inc., and CD62P antibodies (catalog no. 555524) were from BD Biosciences.
4
Palmitate-induced Autophagy and Cell Death
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The chemicals and reagents were from the following sources: ISO (Chengdu Must Bio-Technology Co., Ltd, 52012-29-0), sodium palmitate (Sigma, P9767-5G) that was used to make PA, Dulbecco's modified Eagle's Medium (DMEM, Corning, 10013072), bovine serum albumin (BSA, FDbio Science, FD0030), pentobarbital sodium (Sigma, CAS: 57-33-0), CCK-8 Kit (Beyotime, C0039), Caspase-3 Activity Assay Kit (Solarbio, BC3830), DAPI (Beyotime, P0131-25 ml), Oil Red O Staining Kit (Beyotime, C0157), and chloroquine (CQ, MCE, HY-17598A). The antibodies that we used in the study were from the following sources: LC3B Rabbit mAb (ABclonal, A19665), SQSTM1/p62 Rabbit mAb (ABclonal, A19700), ATG5 Rabbit mAb (ABclonal, A19677), Beclin-1 Rabbit mAb (ABclonal, A7353), anti-mTOR antibody (Abcam, ab32028), phospho-mTOR-S2448 Rabbit mAb (ABclonal, AP0115), β-actin monoclonal antibody (ProteinTech, 66009-1-l g), Goat Anti-Mouse IgG H&L (HRP) (Abcam, ab6789), Goat Anti-Rabbit IgG H&L (HRP) (Abcam, ab6721), and Goat Anti-Rabbit IgG H&L (Alexa Fluor 488, ab150077).
5
DNA Damage Response Protein Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were harvested by 0.25% trypsin digestion and washed twice with ice cold phosphate-buffered saline (PBS). Cell pellets were lysed in lysis buffer (50 mM Tris–HCl, pH 7.5; 150 mM NaCl; 0.5% sodium deoxycholate; 1% NP-40) containing protease inhibitors and phosphatase inhibitors at the concentrations recommended by the manufacturer for 30 min in ice bath. SDS-PAGE and western blotting were performed to analyze the cell lysates according to standard protocols. The following antibodies were used in western blot analyses: phospho-histone H2AX (Ser139) monoclonal antibody (Millipore, USA; Cat. No. 05-636, 1:1000); β-Actin monoclonal antibody (Proteintech, USA; Cat. No. 60008-1-lg, 1:2000); PLK1 monoclonal antibody (Santa Cruz Biotechnology, USA; Cat. No. sc-17783, 1:500); RAD51 polyclonal antibody (Proteintech; Cat. No. 14961-1-AP, 1:2000); BRCA2 polyclonal antibody (Proteintech; Cat. No. 19791-1-AP, 1:500); NBS1 polyclonal antibody (Proteintech; Cat. No. 55025-1-AP, 1:2000); RAD50 polyclonal antibody (Ruiyingbio, China; Cat. No. RLT3963, 1:500); MRE11 polyclonal antibody (Ruiyingbio; Cat. No. RLT2829, 1:1000); BRCA1 polyclonal antibody (Ruiyingbio; Cat. No. RLT0519, 1:500); BCL2 monoclonal antibody (Ruiyingbio; Cat. No. RLT0519, 1:1000).
6
Genistein Modulates ORFV059 Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
OFTu cells mock-infected or infected with OV-SY17 (MOI = 1) and treated with different concentrations of genistein (0 μM, 10 μM, 50 μM, and 100 μM) were collected, washed twice with PBS, and then lysed on ice for 30 min using RIPA lysis buffer (Beyotime, China) containing 1% protease inhibitors. Protein samples were subjected to quantification by a BCA assay (Beyotime, China) prior to Western blot analysis. The equal amounts of protein samples were resolved by SDS-PAGE on 10% gels and then transferred to a PVDF membrane (Millpore, United States). The membranes were blocked in 5% non-fat milk and incubated overnight at 4°C with primary antibodies, such as ORFV059 polyclonal antibody (prepared in our laboratory) or β-actin monoclonal antibody (Proteintech, China). Subsequently, the membranes were incubated with appropriate secondary antibodies. Protein bands were visualized using ECL Western blotting detection reagents. The results were normalized to those for β-actin.
7
HNSCC Cell Lines and Culture Conditions
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HNSCC cell line HPV(-) UMSCC11A, obtained from University of Michigan, HPV(+) cells UMSCC47 and HPV(-) SCC1A were gifts from Ogretmen laboratory, Biochemistry and Molecular Biology, MUSC, Charleston, SC, USA. Cell lines were routinely checked for mycoplasma contamination. SCC1A was grown in Dulbecco’s modified Eagle medium (DMEM-Hyclone) containing 10% fetal bovine serum (FBS) (Atlanta Biologicals), with 100U/ml penicillin and 100 μg/ml streptomycin. UMSCC11A was grown in Dulbecco’s modified Eagle medium (DMEM-Hyclone) containing 10% fetal bovine serum (FBS) with 100U/ml penicillin and 100 μg/ml streptomycin and 1% non-essential amino acids. UMSCC47 was grown in DMEM with 10% FBS, 2 mM l ‐glutamine, 1× non‐essential amino acids solution, and 500 μg/ml gentamicin (Gibco). HuR antibody was obtained from Santa Cruz (cat# sc-5261), EGFR antibody obtained from Cell signaling technology (Cat# 2232S), β-Actin monoclonal antibody was purchased from Proteintech (cat # 60008-1-IG). HPV E6/E7 expressing plasmid p1321 HPV(-)16 E6/E7 was a gift from Peter Howley [16 (link)] (Addgene, catalog # 8641).
8
Fibroblast Regulation by RNF2 and TGF-β1
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Fetal bovine serum was purchased from Biological Industries, and Dulbecco’s modified Eagle’s medium (DMEM) was purchased from HyClone. LipofectamineTM3,000 and Trizol were purchased from Invitrogen. β-actin, RNF2, α-SMA, IL-6, COL 1, and TNF-α primers were purchased from Invitrogen. Recombinant TGF-β1 was purchased from Novoprotein Scientific (Shanghai). RNF2 (Cat:16031-1-AP) polyclonal antibody, β-actin monoclonal antibody, HRP-conjugated goat anti-rabbit IgG (SA00001-2), and goat anti-mouse IgG (SA00001-2) were purchased from Proteintech (China). pEGFP-C2-RNF2 plasmid was stored in our laboratory. PrimeScriptTM IV 1st Strand cDNA Synthesis Mix was bought from TaKaRa. FITC Annexin V apoptosis detection kit I (401007) was purchased from BestBio. Human IL-6 ELISA KIT (ml027379), human TNF-α ELISA KIT (ml077385) and human IL-1β ELISA KIT (ml058059) were purchased from Miblo (Shanghai China). TB Green® Premix Ex TaqTM II was purchased from TaKaRa. COL 1 (WL0088), α-SMA (WL02510) antibodies were purchased from Wanleibio (China). ECL chemiluminescent kit was purchased from NCM Biotech. BeyoClickTM EdU Cell Proliferation Kit was purchased from Beyotime (Cat:C0075S).
9
Cellular Signaling Pathway Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Minimum essential medium (MEM), fetal bovine serum (FBS), sodium pyruvate, penicillin-streptomycin, sodium bicarbonate solution and trypsin-EDTA were purchased from Thermo Fisher Scientific, Inc. (Waltham, MA, USA). RNA Isolater Total RNA Extraction Reagent, HiScript® II Q RT Supermix for qPCR (+gDNA wiper), AceQ® qPCR SYBR®−Green Master Mix, RIPA lysis buffer and a BCA protein quantification kit were obtained from Vazyme Biotech Co., Ltd. (Nanjing, China). The following antibodies were purchased from ProteinTech Group, Inc. (Chicago, IL, USA): JAK2 polyclonal antibody, STAT3 monoclonal antibody, and β-actin monoclonal antibody. Secondary antibodies, including horseradish peroxidase (HRP)-conjugated goat anti-rabbit IgG (H+L) and HRP-conjugated goat anti-mouse IgG (H+L), were obtained from Vazyme Biotech Co., Ltd.
10
Western Blot Analysis of ELOA and SCAF4
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Laemmli sample buffer (Bio-Rad, Hercules, CA, USA) was used to lyse cells and extract total protein. The total protein was resolved by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and then transferred onto polyvinylidene fluoride membrane. The membrane was blocked with 5% skimmed milk powder diluted with Tris-buffered saline Tween-20. Rabbit anti-ELOA polyclonal antibody (NBP1-87040, NOVUS) and anti-SCAF4 polyclonal antibody (PA5-36611, Thermo Fisher) were diluted at 1:1000. β-Actin monoclonal antibody (66009-1-Ig, Proteintech) was diluted at 1:5000. Immunoblotting was imaged by Chemodoc MP (Bio-Rad, Hercules, CA, USA) system.
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!