Sulphorhodamine B (SRB), Neutral Red (NR), (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) (MTT), trichloroacetic acid (TCA), glacial acetic acid, 96% ethanol, hydrochloric acid (HCl), Isopropanol and Tris base were from Sigma Aldrich.
Sulphorhodamine b
Manufactured by Merck Group
Sourced in United States, India, United Kingdom
Sulphorhodamine B is a fluorescent dye commonly used in biological applications. It is a water-soluble, red-orange dye with excitation and emission wavelengths suitable for detection in standard fluorescence assays. Sulphorhodamine B can be used to quantify cellular proteins and measure cell proliferation and cytotoxicity.
Automatically generated - may contain errors
Lab products found in correlation
Trichloroacetic acid, by Merck Group (12 mentions)
Tris base, by Merck Group (5 mentions)
Trypan blue, by Merck Group (5 mentions)
Acetic acid, by Merck Group (5 mentions)
Glacial acetic acid, by Merck Group (3 mentions)
Acetonitrile, by Thermo Fisher Scientific (3 mentions)
Ellipticine, by Merck Group (3 mentions)
Ethanol, by Merck Group (3 mentions)
Hydrochloric acid (hcl), by Merck Group (3 mentions)
Isopropanol, by Merck Group (3 mentions)
Formic acid, by Merck Group (3 mentions)
Hepes, by Merck Group (2 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (2 mentions)
Gallic acid, by Merck Group (2 mentions)
Methanol, by Merck Group (2 mentions)
Penicillin streptomycin, by Merck Group (2 mentions)
L glutamine, by Merck Group (2 mentions)
Resveratrol, by Merck Group (2 mentions)
Trizma base, by Merck Group (2 mentions)
Tryptic soy broth (tsb), by bioMérieux (2 mentions)
33 protocols using sulphorhodamine b
1
Cytotoxicity Assays using Common Reagents
2
Cytotoxicity evaluation of Li, TUD-1, and Li-TUD-1
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All chemicals used in this study (SulphoRhodamine-B (SRB, Tris-HCl, Tricloroacitic acid (TCA)) were purchased from Sigma Chemical Co. (St. Louis, MO, USA). While media and related products were supplemented from Gibco/Life Technologies Co (Carlsbad, CA, USA). Cells Human hepatic carcinoma (HEPG-2), breast (MCF-7) and colon (HCT116) used in the current study were purchased from Vacsera (Giza, Egypt). In preparation to treatment, cells maintained in cell culture media RPMI which is supplemented with 100 µg/mL streptomycin/penicillin and 10% heat-inactivated (FBS) fetal bovine serum. The cytotoxicity of the elements and were tested against HEPG-2, MCF-7, and colon HCT116 tumor cell lines. Confluent 80% cells were exposed to the Li alone, TUD-1 alone and Li-TUD-1 with (0.01, 0.1, 1, 10, 100 µg/mL) for 72 h. SRB evaluation test used to determine the IC50 value (Table 1 ). This value is beneficial and important to evaluate the effective dose for each concentration. The data were analyzed using Sigma Plot version 12.0.
The impact of Li, TUD-1, and Li-TUD-1 on cell viability against cancer cell lines
| Cell type | Cell viability | |||||
|---|---|---|---|---|---|---|
| TUD-1 (%) | Li (%) | Li-TUD-1 | ||||
| 1 | 5 | 10 | 20 | |||
| HEPG2 | 53 | 87.9 | 84.2 ± 1.97 | 89.5 ± 2.40 | 93.42 ± 1.44 | 92.8 ± 1.11 |
| MCF-7 | 56 | 61.4 | 75.9 ± 1.54 | 77.91 ± 3.37 | 80.5 ± 3.9 | 77.6 ± 1.75 |
| HCT116 | 45 | 90.5 | 90.13 ± 1.76 | 92.15 ± 2.46 | 94.04 ± 2.14 | 92.42 ± 1.71 |
3
Clonogenic and Short-term Cytotoxicity Assays
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All short-term survival assays utilized 96-well cell culture plates, into which low passage, exponentially growing cells were seeded at a density of 1,000–4,000 cells per well. The drug was added 24 h postseeding and left for 5 d of continuous exposure. Cellular viability was assessed by CellTitre-Glo luminescence assay (Promega). For clonogenic long-term assays, suspension cells were seeded in six-well plates, coated in Rat tail collagen I. NALM-6WT and NALM-6K700K (3,000 cells per well), NALM-6H662Q cells (3,500 cells per well); K562WT (300 cells per well); K562K700K and K562K700E (650 cells per well). MEL202R625G and MEL202R625G-DEG cells were seeded in standard 6-well plates at 3,500 cells per well and SUM149 cells at 2,000 cells per well. The drug was given 24 h postseeding and to maintain a constant exposure for 14 d and fresh media with inhibitor was replaced every 72 h. For the clonogenic assay, NALM-6 and K562 cell lines were imaged without fixation and quantified on MATLAB vR20018b(9.5.0). For adherent cell lines, the colonies were solubilized with acetic acid and stained with sulphorhodamine B (Sigma-Aldrich), before measuring the optical density at 570–590 nm. Visualization of data was obtained by plotting a line of best fit to 4-parameter nonlinear regression using GraphPad Prism 9 software.
4
Cytotoxicity Evaluation of Bioactive Compounds
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Dimethyl sulfoxide (DMSO, D8418), formaldehyde (61783750001730), and isopropanol (1.94524.2521) were purchased from Merck Ltd., Mumbai, India. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT, M2128), neutral red uptake (NRU, N4638), sulpho rhodamine B (SRB, 230162), antibiotic-antimycotic (Ab/Am, A5955) solution, HEPES (H4034), Propidium iodide (PI, P4170), RNase A (R5000), DCF-DA (D6883), phosphate-buffered saline (PBS, P3813), Rhodamine 123 (R8004), trypsin (93615), ODC (O3001), CATD (C8696), hemoglobin (H2625), sodium bicarbonate (S6297), dihydrofolate reductase (D6566), HYAL (H3884), hyaluronic acid (HA, 924474), cyclooxygenase-2 (COX-2, C0858), methotrexate (MTX, 06563), pepstatin A (P4265), α-difluoro methyl ornithine (DFMO, D193), N-acetyl cysteine (NAC, A7250), celecoxib (32097), zileuton (Z4277), and carvacrol (282197) were purchased from Sigma-Aldrich, Bengaluru, India. Dulbecco's minimal essential media (DMEM) (12100-061), and fetal bovine serum (FBS, 10082147) were procured from Gibco, Thermo Fisher Scientific India Pvt. Ltd., Mumbai, India. LOX-5 (60402) was purchased from Cayman Chemical Company, USA. Sodium/potassium phosphate (RM 257, RM 249) and trichloroacetic acid (RM 6274) were procured from Himedia Ltd, Mumbai, India.
5
Antibody-Based Autophagy and Apoptosis Analysis
6
Colorimetric Cell Viability Assay
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Sulphorhodamine B (SRB) (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) (MTT), trichloroacetic acid (TCA), glacial acetic acid, 96% ethanol, isopropanol and tris base were from Sigma–Aldrich.
7
Cytotoxicity Assessment of V158411
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All cells were obtained from the American Type Culture Collection and cultured in DMEM, RPMI or McCoys 5a containing 10% FCS (Invitrogen). The cytotoxicity of V158411 was determined following exposure of cells in 96 well plates to a 10-point titration for 72 hours. Cell proliferation was determined using sulphorhodamine B (Sigma) staining following protein precipitation with 10% TCA. For cell counts, cells were seeded in 6 well plates and counted following trypsinisation after 72 hours using a haemocytometer with trypan blue staining.
8
Clonogenic and Cell Growth Assays
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For cell growth assays, cells were seeded in 6-well plates (104 per well) and passaged every 3 days until cell numbers were determined. For clonogenic assays, cells were seeded in 6-well plates (400 cells per well) and continuously exposed to ATRi treatment for 14 days, beginning 24 h after seeding. Cells were fixed with 10% trichloroacetic acid and stained with sulphorhodamine B (Sigma-Aldrich). Colonies were counted manually. All cell survival assays were performed at least in triplicate.
9
Antioxidant and Cytotoxicity Assays
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1,1-Diphenyl-2-picrylhydrazyl (DPPH), 2,2′-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS), 2,4,6-tripyridyl-s-triazine (TPTZ), ferrous sulfate (FeSO4.7H2O), aluminium chloride (AlCl3), sodium acetate (C2H3NaO2), sodium carbonate (Na2CO3), potassium persulfate (K2S2O8), potassium chloride (KCl), ferric chloride (FeCl3), sodium nitroprusside {Na2[Fe(CN)5NO].2H2O}, sulfanilic acid (C6H7NO3S), butylated hydroxytoluene (BHT), ascorbic acid, gallic acid, quercetin and catechin were purchased from Sigma-Aldrich (St. Louis, MO, USA). RPMI medium 1640, penicillin, streptomycin, trypsin-EDTA, sulphorhodamine B, trichloroacetic acid, acetic acid, Tris base and all other chemicals used for the cytotoxicity assay were of analytical grade and also purchased from Sigma-Aldrich (St. Louis, MO, USA). Folin Ciocalteu's phenol reagent, vanillin, silica gel precoated aluminium TLC plates (20×20 cm), hydrochloric acid, sulphuric acid, methanol, n-hexane, chloroform, ethanol, toluene, dioxane, acetic acid, diethyl ether, formic acid and sodium carbonate were purchased from Merck Chemical Supplies (Merck KGaA, Darmstadt, Germany). All the other chemicals used including solvents were of analytical grade.
10
SRB Assay for Cell Proliferation
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To measure the effects of analogs on cell proliferation, cells were seeded in 96 well plates at a density 5000 cells per well and after 24 h were treated with serial dilutions of the compounds being tested for an additional 24 h. Following incubation cells were fixed with 10% trichloroacetic acid (TCA, Sigma) for 1 h at 4 °C. Plates were washed five times with distilled water and air-dried. A staining solution comprising 0.4% SRB (sulphorhodamine B, Sigma) in acetic acid was added to each well and after 15 min. plates were washed with 1% acetic acid five times and air-dried. SRB dye was solubilized using a solution of 10 mM buffered Tris Base (pH 10.5). Absorbance was measured at 570 nm using an Epoch spectrophotometer (BioTek, Winooski, VT, USA).
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