The largest database of trusted experimental protocols

Small molecule inhibitors

Manufactured by Selleck Chemicals
Sourced in Germany

Small molecule inhibitors are chemical compounds that can bind to and modulate the activity of specific target proteins. They are designed to interfere with the normal function of these target proteins, which may play a role in disease processes. The core function of small molecule inhibitors is to alter the behavior of the target protein in a controlled manner.

Automatically generated - may contain errors

5 protocols using small molecule inhibitors

1

Preparation and Storage of Mithramycin and PHA-767491

Check if the same lab product or an alternative is used in the 5 most similar protocols
Mithramycin (Tocris Bioscience) was dissolved as a 1 mg/mL stock solution in PBS, and frozen in aliquots. PHA-767491 and other small molecule inhibitors (Selleck Chemicals) were dissolved in DMSO as stock solutions. Small molecule stocks were stored at −80 °C for storage or −20 °C for experimental use, minimizing freeze-thaw cycles. For in vivo experiments, a 30 mg/mL stock solution of PHA-767491 (Adooq Bioscience) was made by dissolving the powder in a solution of 1 mg/10mL D-α-Tocopherol polyethylene 1000 succinate (MilliporeSigma) for oral gavage.
+ Open protocol
+ Expand
2

Cell Line Maintenance and Validation Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols
Small-molecule inhibitors were obtained from SelleckChem. siRNAs were obtained from GE Dharmacon. Cell lines were obtained from American Type Culture Collection (ATCC) and European Collection of Cell Cultures (ECACC) in 2010-2011 and maintained according to the supplier’s instructions as described in (19 (link),47 (link)). The MCF10A CDH1-/- and MCF10A CDH1+/+ isogenic cell lines, were obtained in 2014 from Sigma Aldrich, and maintained according to the supplier’s instructions. At sixth-monthly intervals and prior to storage, the identity of each cell line was confirmed by short tandem repeat (STR) profiling of 10 loci using the GenePrint 10 system (Promega). At monthly intervals, mycoplasma testing of cell cultures was carried out using MycoAlert Mycoplasma Detection Kit (Lonza).
+ Open protocol
+ Expand
3

Naive CD4+ T Cell Activation Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols
Naive CD4+ T cells (CD62L+CD44 or CD62L+CD44FoxP3GFP) were sorted from WT B6, Irf4−/−, or Foxp3gfp reporter mice by a FACSAria flow cytometer. B6 APCs were prepared by depletion of T cells from B6 splenocytes with phycoerythrin–anti-CD3 (clone 2C11; BioLegend) and anti-phycoerythrin microbeads (Miltenyi Biotec, San Diego, CA), followed by brief treatment with 50 μg/ml mitomycin C (Fisher Scientific) before each experiment. For activation of T cells, naïve CD4+ T cells were added at 1×105 cells/well in 96-well round bottom tissue-culture plates (Thermo Fisher Scientific), and stimulated with equal numbers of B6 APCs and 1 μg/ml soluble anti-CD3e mAb (clone 2C11; BioLegend). For some experiments cell cultures were supplemented with various cytokines (PeproTech, Rocky Hill, NJ) and small-molecule inhibitors (Selleck Chemicals). In some cases, naïve CD4+ T cells were labeled with CellTrace™ Violet reagent prior to stimulation. CD4+ T cells cultured for different days were collected and analyzed with flow cytometry, microarray analysis, Immunoblot, and quantitative real-time PCR, and ChIP.
+ Open protocol
+ Expand
4

Culturing Human Melanoma Cell Lines

Check if the same lab product or an alternative is used in the 5 most similar protocols
Human melanoma cell lines (1205Lu, 451Lu, 451Lu‐BR3, WM983B, WM983B‐BR, and WM3734) and HEK293T cells for lentiviral production were provided by Meenhard Herlyn (The Wistar Institute, Philadelphia, USA). Cells were cultured in TU 2% medium (80% MCDB153 basal medium (#P04‐80062, PAN Biotech), 20% Leibovitz's L‐15 medium (#P04‐27055, PAN Biotech) supplemented with 2 mM L‐Glutamin, 1.68 mM CaCl2 and 2% FCS) and were maintained at 37°C in 5% CO2. All cell lines tested negative for Mycoplasma using regularly the PCR Mycoplasma Test Kit I/C (PromoKine #PK‐CA91‐1048, PromoCell GmbH, Heidelberg, Germany). Chemicals were purchased from Sigma‐Aldrich (Munich, Germany), unless otherwise indicated. Small molecule inhibitors were purchased from Selleckchem (Absource, Munich, Germany).
+ Open protocol
+ Expand
5

Cell Line Culture and Authentication

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cell culture medium were obtained from Sigma Aldrich, UK and Life Technologies, UK. Fetal bovine serum (FBS Good) was from PAN Biotech UK Ltd. The HEK293T and LIM1215 cell lines were obtained from the American Tissue Culture Collection, HT115 cells were from Public Health England, DIFI cells were a kind gift from Professor Alberto Bardelli, NCIH508 cells were a kind gift from Dr Anguraj Sadanandam. SNUC4 and SNU1040 cells were obtained from the Korean Cell Line Bank. Cell lines were cultured in medium recommended by the suppliers for up to 3 months at a time, cell line authentication was not performed. Cell lines were tested for mycoplasma using the MycoAlert Mycoplasma Detection Kit (Lonza). Small molecule inhibitors were purchased from Selleck Chemicals. Cetuximab was supplied under a material transfer agreement from Merck.
+ Open protocol
+ Expand

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

Sign up now

Revolutionizing how scientists
search and build protocols!