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A 1155463

Manufactured by Selleck Chemicals
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Sourced in United States, Germany

A-1155463 is a laboratory centrifuge designed for general-purpose sample separation and purification. It features a compact design, digital speed and time controls, and a rotor capable of accommodating various sample tube sizes. The centrifuge is suitable for a range of applications in research and clinical laboratories.

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Lab products found in correlation

Abt 199, by Selleck Chemicals (8 mentions) Abt 737, by Selleck Chemicals (7 mentions) A 1331852, by Selleck Chemicals (5 mentions) Venetoclax, by Selleck Chemicals (5 mentions) S63845, by Selleck Chemicals (4 mentions) Abt 263, by Selleck Chemicals (4 mentions) Cycloheximide (chx), by Merck Group (4 mentions) Oligomycin, by Merck Group (4 mentions) Thapsigargin, by Merck Group (4 mentions) Paclitaxel, by Merck Group (3 mentions) Dimethyl sulfoxide (dmso), by Merck Group (3 mentions) Gemcitabine, by Selleck Chemicals (2 mentions) Mg132, by Thermo Fisher Scientific (2 mentions) Tunicamycin, by Merck Group (2 mentions) Etoposide, by Thermo Fisher Scientific (2 mentions) N acetylcysteine, by Merck Group (2 mentions) Bortezomib, by Thermo Fisher Scientific (2 mentions) 2 deoxyglucose, by Merck Group (2 mentions) Jnk inhibitor 8, by Merck Group (2 mentions) Q vd oph, by Thermo Fisher Scientific (2 mentions)

25 protocols using a 1155463

1

BCL-XL Knockout Cell Viability Assay

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BCL-XL-KO MC38 and Renca cells were generated by transient transfection of Cas9 protein-guide RNA complex into parental cells, followed by single clone section and immunoblot to confirm KO of BCL-XL following the instruction (Synthego). The guide RNA used for BCL-XL KO is 5′-AUACUUUUGUGGAACUCUAU-3′.
Cells were plated in 96-well plates at a density of 3 × 103 to 5 × 103 cells per well and cultured overnight. Cells were treated with increasing concentrations of DT2216, PZ15227, ABT199, a BCL2 specific inhibitor10 (link) (LC laboratories), A1155463, a BCL-XL specific inhibitor38 (link) (Selleck Chemicals, Houston, TX), ABT263, a BCL-2/BCL-XL dual inhibitor9 (link) (Selleck Chemicals) or S63845, a MCL-1 selective inhibitor12 (link) (Selleck Chemicals) for 72 h. Cell viability was measured by tetrazolium-based MTS assay as described previously16 (link). Briefly, MTS reagent (Promega, Madison, WI) was supplemented with phenazine methosulfate (Sigma-Aldrich, St. Louis, MO) at a 20:1 ratio and 20 µl was added to each well and incubated for 4 h at 37 °C. Absorbance was measured at 490 nm using a Synergy Neo2 multimode plate reader (Biotek, Winooski, VT) and cell viability was determined for each well. The data were expressed as the average percent of viable cells and fitted in non-linear regression curves using Prism software (GraphPad Software).
Kolb R., De U., Khan S., Luo Y., Kim M.C., Yu H., Wu C., Mo J., Zhang X., Zhang P., Zhang X., Borcherding N., Koppel D., Fu Y.X., Zheng S.G., Avram D., Zheng G., Zhou D, & Zhang W. (2021). Proteolysis-targeting chimera against BCL-XL destroys tumor-infiltrating regulatory T cells. Nature Communications, 12, 1281.
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2

Evaluating Anti-Apoptotic Proteins in Hematological Malignancies

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All chemicals apart from ABT-199, A1331852, A1155463, A1210477 (Selleck Chemicals, Houston, TX, USA), and S63845 (ApexBio, Taiwan) were from Sigma (Deisenhofen, Germany). Most cell lines used in this study were obtained from Deutsche Sammlung von Mikroorganismen und Zellkulturen (DSMZ; Braunschweig, Germany) except Pfeiffer and SUDHL2 cells (American Type Culture Collection; Manassas, VA, USA), OCI-LY10 (Sandeep Dave, Duke University, Durham, NC, USA), MedB116 (link) (Peter Moeller, University of Ulm, Ulm, Germany) and Karpas-110617 (link) (Abraham Karpas, University of Cambridge, Cambridge, UK). All cell lines were authenticated by short tandem repeat profiling and routinely tested for mycoplasma contamination. Primary patient-derived samples were obtained from patients attending the University Hospital of Leicester, UK. Local ethical approval (Leicestershire, Northamptonshire and Rutland REC06/Q2501/122) and patients’ consent were obtained through the Haematological Tissue Bank of the Ernest and Helen Scott Haematological Research Institute, Leicester, UK. Peripheral blood mononuclear cells were isolated from the blood of patients presenting in leukemic phase and the CellTiterGlo assay (Promega, Mannheim, Germany) was used to assess these cells’ viability.
Smith V.M., Dietz A., Henz K., Bruecher D., Jackson R., Kowald L., van Wijk S.J., Jayne S., Macip S., Fulda S., Dyer M.J, & Vogler M. (2020). Specific interactions of BCL-2 family proteins mediate sensitivity to BH3-mimetics in diffuse large B-cell lymphoma. Haematologica, 105(8), 2150-2163.
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3

Synthesis and Utilization of DT2216 Compound

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DT2216 was synthesized in Dr. Guangrong Zheng's laboratory (University of Florida, Gainesville, FL) according to the previously described protocol (20 (link)). Gemcitabine (catalog No. S1714), 5FU (catalog No. S1209), niraparib (catalog No. S2741), A1155463 (catalog No. S7800), ABT199 (catalog No. S8048), S63845 (catalog No. S8383), and ABT263 (catalog No. S1001) were purchased from SelleckChem. All the compounds were dissolved in DMSO at 10 mmol/L stock solution for in vitro assays.
Thummuri D., Khan S., Underwood P.W., Zhang P., Wiegand J., Zhang X., Budamagunta V., Sobh A., Tagmount A., Loguinov A., Riner A.N., Akki A.S., Williamson E., Hromas R., Vulpe C.D., Zheng G., Trevino J.G, & Zhou D. (2021). Overcoming Gemcitabine Resistance in Pancreatic Cancer Using the BCL-XL–Specific Degrader DT2216. Molecular Cancer Therapeutics, 21(1), 184-192.
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4

Evaluation of Bioactive Compounds in Cell Screening

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Camptothecin (Cat# S1288), vinblastine (Cat# S1248), pimasertib (Cat# S1457), trametinib (Cat# S2673), ABT-737 (Cat# S1002), A-1155463 (Cat# S7800), and nutlin-3 (Cat# S1061) were purchased from Selleck Chemicals (Houston, TX). Bortezomib (Cat# NC0587961), etoposide (Cat# ICN19391825), MG-132 (Cat# 17-485), and Q-VD-OPh (Cat# OPH00101M) were purchased from Thermo Fisher Scientific. N-acetylcysteine (Cat# A8199), thapsigargin (Cat# T9033), tunicamycin (Cat# T7765), paclitaxel (Cat# T7191), JNK Inhibitor VIII (Cat# 420135), 2-deoxyglucose (Cat# D8375), oligomycin (Cat# O4876), and cycloheximide (Cat# C7698) were obtained from Sigma-Aldrich (St. Louis, MO). S63845 (Cat#21131) was obtained from Cayman Chemical (Ann Arbor, MI). Staurosporine (Cat# A8192) was obtained from ApexBio (Houston, TX). Erastin was the kind gift of Brent Stockwell (Columbia University). Erastin2 (compound 35MEW28 in Dixon et al., [2014 (link)]) and ML162 (CAS: 1035072-16-2) were synthesized by Acme Bioscience (Palo Alto, CA). Chemical screening was conducted as described below; the library of 261 bioactive compounds was obtained from Selleck Chemicals (Cat# L2000).
Inde Z., Forcina G.C., Denton K, & Dixon S.J. (2020). Kinetic Heterogeneity of Cancer Cell Fractional Killing. Cell reports, 32(1), 107845.
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5

Quantifying Cell Death Pathways

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FL-P or FL-D cells were plated in 24-well plates (105 cells in 500 µl Hoxb8 progenitor or B cell medium) and treated with etoposide (Sigma Aldrich) or the inhibitors ABT-737, ABT-199, A-1155463 (Selleckchem) or S53845 (ApexBio) as indicated. For analysis of cell death induced by factor withdrawal, cells were washed and plated without FLT3L (FL-P-cells) or IL-7 (FL-D-cells). At various time points, cells were collected in PBS/4% FCS. Propidium iodide (Sigma Aldrich) was added immediately prior to analysis by flow cytometry (FACS-Calibur, Becton Dickinson). In some experiments, annexin V-Propidium iodide staining was used for quantification of apoptosis. Cell were washed with annexin V-binding buffer (eBioscience) and stained with annexin V-FITC (1:20, BD Pharmingen) and PI (5 µg/ml) for 20 min at 4 °C followed by flow cytometry analysis (FACS Calibur). For staining of active caspase-3, cells were fixed in 2% paraformaldehyde and permeabilized with 0.5% saponin (Sigma-Aldrich). Cells were incubated with anti-active caspase-3 (BD Pharmingen) in PBS/0.5%BSA/0.5% saponin for 30 min, stained with anti-rabbit-Alexa-Fluor488 (Dianova GmbH, Hamburg, Germany) for 30 min and analyzed by flow cytometry (FACS Calibur). Routinely, three biological replicates were performed. Further replicates were done depending on the statistical distribution of the values.
Kurschat C., Metz A., Kirschnek S, & Häcker G. (2021). Importance of Bcl-2-family proteins in murine hematopoietic progenitor and early B cells. Cell Death & Disease, 12(8), 784.
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6

Compound Screening for Cellular Assays

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ABT-199, ABT-263, ABT-737, WEHI-539, A-1331852, A-1155463, obatoclax, and gemcitabine were purchased from Selleck Chemicals (Houston, TX, USA), Saliphenylhalamide (SaliPhe) was synthesized as described previously [24 (link)]. 10 mM solutions of the compounds were prepared in 100% dimethyl sulfoxide (Sigma-Aldrich, St. Louis, MO, USA) and stored at −80 °C. Lyophilized lipopolysaccharide (LPS, 10 mg/mL) was purchased from Sigma-Aldrich (St. Louis, MO, USA). Plasmid pEGFP was from Clontech (Mountain View, CA, USA) (cat #: HLP309). Hoechst 33342 (20 mM solution; cat #: 62249) and ATP (10 mM solution; cat #: PV3227) were from Thermo Fisher Scientific (Waltham, MA, USA). Genomic RNA was isolated from influenza A/WSN/1933 strain as described previously [11 (link)].
Bulanova D., Ianevski A., Bugai A., Akimov Y., Kuivanen S., Paavilainen H., Kakkola L., Nandania J., Turunen L., Ohman T., Ala-Hongisto H., Pesonen H.M., Kuisma M.S., Honkimaa A., Walton E.L., Oksenych V., Lorey M.B., Guschin D., Shim J., Kim J., Than T.T., Chang S.Y., Hukkanen V., Kulesskiy E., Marjomaki V.S., Julkunen I., Nyman T.A., Matikainen S., Saarela J.S., Sane F., Hober D., Gabriel G., De Brabander J.K., Martikainen M., Windisch M.P., Min J.Y., Bruzzone R., Aittokallio T., Vähä-Koskela M., Vapalahti O., Pulk A., Velagapudi V, & Kainov D.E. (2017). Antiviral Properties of Chemical Inhibitors of Cellular Anti-Apoptotic Bcl-2 Proteins. Viruses, 9(10), 271.
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7

Mitochondrial and Cellular Function Assays

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Oligomycin (Cat# O4876), Antimycin A (Cat# A8674), nigericin (Cat# N7143), cycloheximide (Cat# C4859), rotenone (Cat# R8875), ionomycin (Cat# 13909), thapsigargin (Cat# T9033), staurosporine (Cat# S6942), Actinomycin D (Cat# A9415) were purchased from Sigma Aldrich. Piercidin (Cat# SC‐202287) was purchased from Chemcruz. Venetoclax (Cat# S8048) and A‐1155463 (S7800) were purchased from Selleckchem. Fura2 (Cat# F1200), TMRM (Cat# T668) and MitoSox (Cat# M36008) were purchased from Invitrogen. Calcium‐Green‐5N (Cat# C3737) was purchased from life technologies.
Patron M., Tarasenko D., Nolte H., Kroczek L., Ghosh M., Ohba Y., Lasarzewski Y., Ahmadi Z.A., Cabrera‐Orefice A., Eyiama A., Kellermann T., Rugarli E.I., Brandt U., Meinecke M, & Langer T. (2022). Regulation of mitochondrial proteostasis by the proton gradient. The EMBO Journal, 41(16), e110476.
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8

Ovarian Cancer Sensitivity to Carboplatin

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In order to detect the sensitivity of ovarian cancer cells (cultured in adipocyte-conditioned medium and adipocyte co-culture, and regulated by multiple small molecular inhibitors, agonists and neutralizing antibodies in vitro) to carboplatin chemotherapy, SKOV3 cells (wild-type or gene-knockdown type) were added into the 96-well plate (Corning, Inc.) at the density of 6,000 cells per well, and the medium was removed 24 h after cells reached equilibrium. Subsequently, DMEM containing various pathway regulators (20 µg/ml cANGPTL4, 48 µg/ml anti-cANGPTL4, 50 µM verapamil (Aladdin) and 50 nM A-1155463 (Selleck Chemicals), chemotherapeutics (carboplatin) or adipocyte-conditioned medium was added to the cells; alternatively, the Transwell upper chamber (Corning, Inc.) incubated with adipocytes was combined with the SKOV3 cells for 48 h of co-culture. All drugs were prepared at appropriate concentrations using DMSO, and equivalent amount of DMSO was present in each well of medium. Subsequently, the CellTiter-Glo reagent (Promega Corporation) was added into the cell medium according to the manufacturer's protocol, and the luminance value was measured using the BioTek Synergy 96-well microplate reader. All experiments were repeated three times.
Zhou S., Wang R, & Xiao H. (2020). Adipocytes induce the resistance of ovarian cancer to carboplatin through ANGPTL4. Oncology Reports, 44(3), 927-938.
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9

Apoptosis Induction in Cell Lines

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For cell death experiments, cells were seeded in six-well plates at a confluence of 1,000,000 cells/well. The next day, the cells were treated with staurosporine (1 μM), thapsigargin (2 μM), or selective BH3-mimetic inhibitors of the Bcl-2 family of proteins (venetoclax as Bcl-2 inhibitor and A1155463 as Bcl-XL inhibitor) (1 μM) (Sellekchem) for 12 h. Subsequently, cells were collected and pelleted by centrifugation and incubated with Annexin V-FITC (Life Technologies, Carlsbad, CA, USA, V13245) and 7-AAD (Becton Dickinson, Franklin Lakes, NJ, USA, 555815). Cell suspensions were analyzed with an Attune Acoustic Focusing Flow Cytometer (Applied Biosystems, Waltham, MA, USA). Cell death by apoptosis was scored by quantifying the population of Annexin V-FITC-positive cells using the FlowJo version 10 software. Data are plotted as the ∆ apoptotic fraction, which is calculated as the difference between the percentage of apoptotic cells in the compound-treated condition and the percentage of apoptotic cells in the vehicle-treated condition.
Seitaj B., Maull F., Zhang L., Wüllner V., Wolf C., Schippers P., La Rovere R., Distler U., Tenzer S., Parys J.B., Bultynck G, & Methner A. (2020). Transmembrane BAX Inhibitor-1 Motif Containing Protein 5 (TMBIM5) Sustains Mitochondrial Structure, Shape, and Function by Impacting the Mitochondrial Protein Synthesis Machinery. Cells, 9(10), 2147.
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10

Protein Stability and Cell Viability Assays

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Protein stability was followed by treatment with cycloheximide (Sigma; Cat# C4859) or emetine (Sigma; Cat# E2375) at a concentration of 100 μg × ml−1 over the course of 8 h. At indicated time points, cells were collected and lysed according to the protocol above. Cell viability was assessed after treatment with actinomycin D (Sigma, Cat# A9415; 1 μM), venetoclax (Selleckchem; Cat# S8048; 1 μM), A‐1155463 (Selleckchem; Cat# S7800; 1 μM), staurosporine (Sigma, Cat# S6942; 1 μM) or thapsigargin (Sigma; Cat# T9033; 2 μM) for 16 h. ATP synthase was inhibited by oligomycin treatment (Sigma, Cat# O4876; 10 μM) for 18 h.
Patron M., Tarasenko D., Nolte H., Kroczek L., Ghosh M., Ohba Y., Lasarzewski Y., Ahmadi Z.A., Cabrera‐Orefice A., Eyiama A., Kellermann T., Rugarli E.I., Brandt U., Meinecke M, & Langer T. (2022). Regulation of mitochondrial proteostasis by the proton gradient. The EMBO Journal, 41(16), e110476.
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