Invasion assay was measured by counting the THLE-2 invaded cells after overexpression with empty N-Terminal p3XFLAG-CMV plasmid (Sigma-Aldrich, Saint Louis, MO, USA) or with plasmid containing BRAF WT and BRAF V600E. Corning BioCoat™ Matrigel Invasion Chamber (Corning, NY, USA) with 8 μm pores inserts was used to perform the analysis according to manufacturer instructions. After 24 h from transfections THLE-2 cells were seeded in the number of 0.5 × 105 in serum-free LHC-8 medium and incubated for 24 h under standard conditions. Non-invasive cells from the upper surface of the membrane were removed with cotton swabs. The invading cells were fixed and stained using Differential Quick III Stain Kit (Polysciences, Inc., Warrington, PA, USA), and photographed under the light microscope (Nikon Eclipse E200, Tokyo, Japan). The number of cells was calculated using ImageJ software and presented as a bar chart.
Differential quick 3 stain kit
Manufactured by Polysciences
Sourced in United States
The Differential Quick III Stain Kit is a laboratory product designed for the rapid staining of blood smears. It provides a simple and efficient method for differentiating and identifying various types of blood cells. The kit includes the necessary staining solutions to perform the staining procedure.
Automatically generated - may contain errors
Lab products found in correlation
N terminal p3xflag cmv plasmid, by Merck Group (1 mentions)
Biocoat matrigel invasion chamber, by Corning (1 mentions)
Hypoxanthine, by Merck Group (1 mentions)
Gentamicin, by Thermo Fisher Scientific (1 mentions)
Chloroquine, by Merck Group (1 mentions)
L glutamine, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
Sodium bicarbonate, by Merck Group (1 mentions)
Atcc pra 405d, by American Type Culture Collection (1 mentions)
2 protocols using differential quick 3 stain kit
1
BRAF-Driven Cell Invasion Assay
2
Antimalarial Drug Screening Protocol
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Chloroquine, sodium bicarbonate, and hypoxanthine were purchased from Sigma-Aldrich (St. Louis, MO, USA). Gentamicin, SYTOX green solution, AlbuMAX I Lipid-Rich bovine serum albumin, and RPMI 1640 medium supplemented with 25 mM HEPES and L-glutamine were purchased from Gibco (Life Technologies Corporation, Grand Island, NY, USA). The Differential Quick III Stain Kit was purchased from Polysciences Inc. (Valley Road, Warrington, PA, USA). CQ-sensitive strain P. falciparum (3D7) (American Type Culture Collection, ATCC PRA-405D) and Vero cells (CRL-1587) were obtained from the American Type Culture Collection (ATCC) (Manassas, VA, USA). A multiple antimalarial drug-resistant strain, P. falciparum K1 (MRA-159), and a CQ-susceptible strain, P. berghei NK65 (MRA-268), were obtained from BEI Resources (Manassas, VA, USA) [11 (link)].
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