Vero E6, Vero-ACE2-TMPRSS2, Huh-7, Vero 81, and 293T cells were maintained in Dulbecco’s minimal essential medium (DMEM) containing 10% fetal bovine serum (FBS) and antibiotics [chlortetracycline (25 µg/mL), penicillin (250 U/mL), and streptomycin (250 µg/mL)]. Calu-3 2B4 cells were grown in DMEM supplemented with 20% FBS and antibiotics. MA-SARS-CoV-2 (SARS2-N501YMA30) (56 (link)) and MA-MERS-CoV (57 (link)) were propagated in Calu-3 2B4 and Huh-7, respectively. SARS-CoV-2 XBB.1.16 omicron variant (isolate hCoV-19/USA/CA-Stanford-139_S23/2023) was obtained from BEI. Lentivirus-based pseudoviruses expressing SARS-CoV-2 Wuhan-Hu-1 strain were generated with the second-generation lentiviral packaging plasmid psPAX2 (Addgene, Watertown, MA), a reporter plasmid pUCGFP-Luc (Addgene), and pAbVec-SARS2-S (42 (link)). Both plasmids expressing hACE2 or TMPRSS2 were obtained from Origen (Rockville, MD). MDL28170 (calpain and cathepsin B inhibitor) and Nafamostat (TMPRSS2 inhibitor) were purchased from Sigma-Aldrich (St Louis, MO). SARS-CoV-2 3CLpro inhibitors, 5c/d and 11c/d were recently reported from our labs (41 (link)). Polybrene was purchased from Santa Cruze Biotechnology (Santa Cruz, CA).
Mdl28170
Manufactured by Merck Group
Sourced in United States, Germany
MDL28170 is a laboratory product manufactured by Merck Group. It is a heterocyclic organic compound used in research and development applications. The core function of MDL28170 is to serve as a chemical tool for scientific investigations, without further interpretation of its intended use.
Automatically generated - may contain errors
Lab products found in correlation
Camostat mesylate, by Merck Group (2 mentions)
Zvad fmk, by Bachem (2 mentions)
Nafamostat, by Merck Group (1 mentions)
Pucgfp luc, by Addgene (1 mentions)
Pspax2, by Addgene (1 mentions)
Streptozotocin (stz), by Merck Group (1 mentions)
Amiloride, by Merck Group (1 mentions)
Anti egfp, by Thermo Fisher Scientific (1 mentions)
A6455, by Thermo Fisher Scientific (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
Anti β tubulin antibody, by Merck Group (1 mentions)
Dynabeads protein g, by Thermo Fisher Scientific (1 mentions)
Monobromobimane, by Merck Group (1 mentions)
Chloroquine, by Merck Group (1 mentions)
Anti phosphotyrosine antibody clone 4g10, by Merck Group (1 mentions)
Antibodies against β actin, by Santa Cruz Biotechnology (1 mentions)
Caspase 8, by Cell Signaling Technology (1 mentions)
Raf 1, by Santa Cruz Biotechnology (1 mentions)
Geldanamycin, by Sangon (1 mentions)
Poly adp ribose polymerase, by Cell Signaling Technology (1 mentions)
21 protocols using mdl28170
1
SARS-CoV-2 Variant Cell-Based Assays
2
Diabetic Mouse Model Treatment
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The study was approved by the Animal Care and Use Committee of Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China. Thirty-eight-week-old male specific-pathogen-free C57BL/6J mice obtained from the Laboratory Animal Center of Tongji Medical College, Huazhong University of Science and Technology, were randomly divided into three groups: nondiabetic control (n = 10), diabetic mice treated with vehicle (a 1:1 solution of DMSO [dimethyl sulfoxide]: PBS [phosphate buffer saline]; n = 10), and diabetic mice treated with MDL28170 (Sigma-Aldrich, M6690, St. Louis, MO, USA; dissolved in a 1:1 solution of DMSO: PBS at a concentration of 2 g l−1; n = 10). All mice were kept under 12-h light/dark conditions. Type 1 diabetes was induced by intraperitoneal injection of streptozotocin (Sigma-Aldrich, S0130) at 60 mg kg−1 body weight for 5 consecutive days. Blood glucose levels were measured 1 week after the last injection and mice with a fasting glucose concentration ≥16.7 mmol l−1 were considered as diabetic mice. The fasting blood glucose and body weight of all mice were then measured once every 4 weeks. Twelve weeks later, diabetic mice were injected intraperitoneally with 20 mg kg−1 body weight MDL28170 or vehicle daily for 4 weeks according to the grouping.21 (link)
3
Studying C2R10 Virus Inhibition
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Rhileki cells were seeded in 24-well plates at a density of 3 × 105 cells per well overnight. On the day of infection, cells were washed once with PBS and treated with medium, 50 μM MDL28170 (Sigma-Aldrich, USA), or 10 μM E64d (Tocris Bioscience, UK) for 1 h at 37°C. After pretreatment, cells were spin inoculated in medium containing either of the two compounds and C2R10 viruses at an MOI of 0.003 by centrifugation at 1,500 × g for 1 h at 4°C. Thereafter, the inoculum was removed, and cells were incubated with medium containing the corresponding chemical for 5 h before replenishment of medium without chemicals and incubation at 33°C for 72 h.
4
Immunostaining and Microscopy Techniques
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Dulbecco’s phosphate buffered saline (PBS) (number D5652), bovine serum albumin (BSA) (number A9647), amiloride (number A-7410), and MDL28170 (number M6690) were provided by Sigma-Aldrich, St. Louis, MO, USA. The polyclonal rabbit primary antibodies were anti-EGFP (Molecular Probes, A6455, Thermo Fisher, Waltham, MA, USA). The monoclonal mouse antibodies used were anti-GFP (B-2) (sc-9996, Santa-Cruz Biotechnology, Dallas, TX, USA), 2B4, and 1C2 (IGBMC antibody platform, Illkirch, France). The other antibodies were secondary anti-rabbit (sc-2004, Santa-Cruz Biotechnology, Dallas, TX, USA) and anti-mouse (NA931, GE Healthcare, Bath, UK) antibodies conjugated to horseradish peroxidase; for immunostaining: anti-rabbit and anti-mouse antibodies conjugated to Alexa-488 (CAR-A21441), to Alexa-594 (GAM- A21422, DAM-A31570) from Molecular Probes (Thermo Fisher, Waltham, MA, USA). For immunoelectron microscopy, anti-rabbit antibodies conjugated with 15 nm gold beads (Auroprobe TM/EM, LRPN422V/AA/G15, Auroprobe, New Delhi, India), and anti-mouse antibodies conjugated with 10 nm gold beads (G7777, Sigma-Aldrich, St. Louis, MO, USA) were used. The vectors used for HTT and mHTT expression were pEGFP-C2-tr/HTT-17Q and pEGFP-C2-tr/HTT-142Q, generated at IGBMC (Strasbourg, France).
5
Immunoprecipitation and Calpain Activity Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Primary antibodies against calpain-1 and 2 and TrkB were purchased from Cell Signaling Technology, Danvers, MA, USA and anti-β-tubulin antibody was from Sigma, St. Louis, MO, USA. Anti-calpain-1 antibody used for immunoprecipitation of calpain-1 was procured from Millipore. Purified anti-β-Amyloid, 1–16 antibody (6E10) (Cat. No. 803003; RRID: AB_2564652) was obtained from BioLegend Inc., USA. Immunoprecipitation of calpain-1 and calpain-2 was performed using Dynabeads Protein G and Protein A, respectively (Life Technologies, CA, USA) and calpain activity assay was performed using a fluorogenic substrate N-succinyl-Leu-Leu-Val-Tyr-7-amino-4-methylcoumarin (Suc-LLVY-AMC; Farmingdale, NY, USA). Calpain inhibitor, MDL 28170 was procured from Sigma-Aldrich. All other chemicals and reagents used were of analytical grade and obtained from either Sigma-Aldrich or Merck.
6
Inhibition of Hsp90 and Proteasome in Cancer Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Y‐632 was synthesized at the Shanghai Institute of Materia Medica, Chinese Academy of Sciences (Shanghai, China). Imatinib mesylate was obtained from Selleck Chemicals (Houston, TX, USA). Geldanamycin was obtained from Sangon Biotech (Shanghai, China). MG132, proteasome inhibitor I (PS341), chloroquine, MDL28170, monobromobimane, SRB, MTT, and antibodies against β‐tubulin and insulin‐like growth factor 1 receptor were purchased from Sigma‐Aldrich (St. Louis, MO, USA). Antibodies against human epidermal growth factor receptor‐2, GAPDH, EGFR, c‐Met, Akt, phospho‐Akt (Ser473), Cdk4, Cdk6, p85, poly‐(ADP‐ribose) polymerase, caspase 8, caspase 9, Hop, Cdc37, Erk1/2, and phospho‐Erk1/2 (Thr202/Tyr204) were purchased from Cell Signaling Technology (Beverly, MA, USA). Antibodies against β‐actin, Raf‐1, p23, and caspase 3 were from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Antibodies against Hsp90 and Bcr‐Abl were from Abcam (Cambridge, UK). Anti‐Hsp70 antibody was from Enzo Life Sciences (Farmingdale, NY, USA). Antiphosphotyrosine antibody (clone 4G10) was obtained from Millipore (Billerica, MA, USA).
7
Neuroprotective Effects of Memantine and MDL-28170
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The NMDAR antagonist MEMantine (MEM, 20 mg/kg, Sigma, St Louise, MO, USA) [24 (link)] was intraperitoneally (i.p.) administered to the mice before anesthesia and then once daily for subsequent 5 consecutive days. The calpain inhibitor, MDL-28170 (20 mg/kg, Sigma, St Louise, MO, USA) [25 (link)], was administered to the mice by i.p. before anesthesia and then once daily for the subsequent 5 consecutive days. The selected doses are based on previous studies demonstrating MEMantine and MDL-28170 confers neuroprotective effects [24 (link), 25 (link)].
8
MERS-S Activation Protease Inhibition
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
9
Studying C2R10 Virus Inhibition
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Rhileki cells were seeded in 24-well plates at a density of 3 × 105 cells per well overnight. On the day of infection, cells were washed once with PBS and treated with medium, 50 μM MDL28170 (Sigma-Aldrich, USA), or 10 μM E64d (Tocris Bioscience, UK) for 1 h at 37°C. After pretreatment, cells were spin inoculated in medium containing either of the two compounds and C2R10 viruses at an MOI of 0.003 by centrifugation at 1,500 × g for 1 h at 4°C. Thereafter, the inoculum was removed, and cells were incubated with medium containing the corresponding chemical for 5 h before replenishment of medium without chemicals and incubation at 33°C for 72 h.
10
Decoy peptide for Ser16 phosphorylation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The decoy peptide (RAE16TIEMPQ) was derived from the PLB protein sequence surrounding the Ser16 phosphorylation site. To facilitate uptake into cells, the decoy peptide was conjugated to the cell penetrating peptide TAT (YGRKKRRQRRR). The peptides used in this study were ΨPLB-SE and RAETIEMPQ. The peptides (purity, >95%; AnyGen, Gwangju, Korea) were resuspended in double-distilled water at a stock concentration of 3 mM. RASMC and HCSMC were treated with the peptides at a final concentration of 3 μM for 1 h. The calcium ionophore A23187, the calpain I and II inhibitor MDL28170, cycloheximide were purchased from Sigma—Aldrich (St. Louis, MO, USA).
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!