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Noxa 114c307.1

Manufactured by Thermo Fisher Scientific
Sourced in United States

The Noxa (114C307.1) is a lab equipment product manufactured by Thermo Fisher Scientific. It is a specialized device designed for use in laboratory settings. The core function of this product is to perform specific tasks related to laboratory procedures, but a detailed description cannot be provided while maintaining an unbiased and factual approach.

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7 protocols using noxa 114c307.1

1

Immunoprecipitation and Western Blot Analysis

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Antibodies were purchased as follows: BCL-2 (clone 100) from Sigma-Aldrich (St. Louis, MO) and Cell Signaling (GAPDH, BIM (C34C5), BCL-XL (54H6), and Cleaved PARP (D64E10) from Cell Signaling Technology (Danvers, MA); Noxa (114C307.1) from Thermo Fisher Scientific (Waltham, MA); MCL-1 from Enzo Life Sciences (Farmingdale, NY); BAX (N-20) from Santa Cruz Biotechnology (Santa Cruz, CA). Immunoprecipitation and Western blots were performed as described in (19 (link)). For Sup. Fig. 12, 500ug of each lysate were incubated with BIM antibody (500 ng; Cell Signaling Technology, cat# 2933S), or rabbit IgG (500 ng; Santa Cruz Biotechnology, cat# sc-2027). Following the addition of 25uL of 1:1 PBS: pre-washed Protein A beads to the antibody/lysate mix, samples were incubated with rotating motion overnight. Equal amounts of extracts (5% of immunoprecipitated protein) were prepared in parallel.
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2

Apoptosis Induction Pathway Analysis

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Fenretinide, ABT-263, and Q-VD-OPh were purchased from ApexBio Tech (Houston, TX). Antibodies for BIM, BAX, BAK, BCL-XL, Cleaved-PARP (Asp214), Cleaved-Caspase3, GAPDH (D16H11), HRP-linked Anti-Rabbit IgG, HRP-linked anti-mouse IgG, and Anti-Rabbit IgG conformation-specific antibodies were from Cell Signaling Technology (Beverly, CA); NOXA (114C307.1) was from Thermo Fisher Scientific (Waltham, MA); MCL-1 (ADI-AAP-240-F) was from Enzo Life Sciences (Farmingdale, NY); BCL-2 (100) was from Sigma-Aldrich. BAK antibodies for immunoprecipitation were purchased from Sigma-Aldrich (06–536 and AM03). ECL2 Western blotting substrate was purchased from Thermo Fisher Scientific (Rockland, IL). Annexin V-FITC and Annexin V-APC were purchased from Biolegend (San Diego, CA) and Thermo Fisher Scientific, respectively. Propidium Iodide was purchased from Sigma-Aldrich.
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3

Antibody Acquisition and Small Molecule Procurement

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Antibodies were purchased as follows: BIM (C34C5), BCL-XL (54H6), CREB (48H2) and cleaved caspase-3 (5A1E) from Cell Signaling Technology (Danvers, MA, USA); NOXA (114C307.1) from Thermo Fisher Scientific (Waltham, MA, USA); ATF-3 (C-19) and MCL-1 (S-19) from Santa Cruz Biotechnology (Santa Cruz, CA, USA); KLF4 (56CT5.1.6) from Bioss, Inc. (Woburn, MA, USA); α-tubulin (DM1A) and β-actin (AC-74) from Sigma-Aldrich (Tokyo, Japan). APTO-253 was purchased from MedChemExpress, LLC (Monmouth Junction, NJ, USA). Q-VD-OPh, Doxorubicin, etoposide, SP600125, SB203580, PD184352, and U0126 were purchased from Calbiochem (San Diego, CA, USA). Hydroxychloroquine (HCQ), Necrostatin-1 and Ferrostatin-1 were purchased from Cayman Chemical (Ann Arbor, MI, USA).
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4

Reagents and Antibodies for Cell Signaling

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Reagents were purchased as follows: cisplatin from Santa Cruz Biotechnology (Santa Cruz, CA, USA); etoposide and SP600125 from LC Laboratories (Woburn, MA, USA); SB203580, Roscovitine, LY294002, and CHIR99021 from Sigma-Aldrich; Cycloheximide and MG-132 from Millipore (Billerica, MA, USA); Phos-tag acrylamide from Wako Chemicals USA (Richmond, VA). PD184352 was kindly provided by Steven Grant (Virginia Commonwealth University), which was chemically synthesized in house based on the published structure of the drug. cisplatin was dissolved in PBS [30 (link)] and other reagents were dissolved in dimethyl sulfoxide. The following antibodies were used: Cleaved PARP (D64E10), Cleaved Caspase-3 (5A1E), BCL-XL, BIM, phospho-S64 MCL-1, COX IV (3E11), β-Actin (13E5) and GAPDH from Cell Signaling Technology (Danvers, MA, USA); Noxa (114C307.1) from Thermo Fisher Scientific (Waltham, MA, USA); MCL-1 from Enzo Life Sciences (Farmingdale, NY, USA) and Rockland Immunochemicals (Gilbertville, PA, USA); BAK (06-536) from Millipore; BAX (N-20), alpha-Tubulin (sc-8035), Myc (9E10) and CDK1 (sc-54) from Santa Cruz Biotechnology; CDK2 from BD Biosciences; HA (3F10) from Roche (Indianapolis, IN, USA). [γ-32P]ATP was purchased from PerkinElmer (Kanagawa, Japan). Histone H1, recombinant CDK2/Cyclin A, and CDK2/Cyclin E were purchased from SignalChem (Richmond, BC, Canada).
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5

Apoptosis Protein Expression Analysis

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Cycloheximide and MG132 were purchased from Millipore (Billerica, MA, USA). Antibodies purchased were as follows: BIM (C34C5), BCL-XL (54H6), Cleaved PARP (D64E10) and Cleaved Caspase-3 (5A1E) from Cell Signaling Technology (Danvers, MA, USA); Noxa (114C307.1) from Thermo Fisher Scientific (Waltham, MA, USA); BAX (6A7) from Sigma (St. Louis, MO, USA); MCL-1 from Enzo Life Sciences (Farmingdale, NY, USA); BAX (N-20) and alpha-Tubulin (sc-8035) from Santa Cruz Biotechnology (Santa Cruz, CA, USA); BAK (06–536) and BAK (Ab-1) from Millipore; and Ubiquitin (VU-1) from Life Sensors (Malvern, PA, USA).
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6

Antibody Acquisition for Protein Analysis

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Antibodies were purchased as follows: BIM (C34C5), BCL-XL (54H6), PARP (#9542), Cleaved Caspase-3 (5A1E), DNMT1 (D63A6), DNMT3A (D23G1), and DNMT3B (E8A8A) from Cell Signaling Technology (Danvers, MA, USA); NOXA (114C307.1) from Thermo Fisher Scientific (Waltham, MA, USA); MCL1 (S-19), p21 (F-5) and ATF3 (C-19) from Santa Cruz Biotechnology (Santa Cruz, CA, USA); Anti-KLF4 (56CT5.1.6) from Bioss Inc. (Woburn, MA, USA); β-actin (AC-74) from Sigma-Aldrich (Tokyo, Japan); p27 (554069) and p16 (554079) from BD Pharmingen (San Jose, CA, USA); and ATF4 (739441) from R&D Systems (Minneapolis, MN, USA).
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7

Investigating Cisplatin-Induced Cell Stress

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Cisplatin and SP600125 were purchased from ApexBio (Houston, TX, USA). SB203580 and PD184352 were purchased from LC Laboratories (Woburn, MA, USA). Cisplatin was dissolved in PBS and other reagents were dissolved in dimethyl sulfoxide (Hall et al., 2014). The following antibodies were used: cleaved‐PARP (D64E10), ATF4, CHOP and GAPDH from Cell Signaling Technology (Danvers, MA, USA); Noxa (114C307.1) from Thermo Fisher Scientific (Waltham, MA, USA); MCL‐1 from Enzo Life Sciences (Farmingdale, NY, USA); ATF3, ATF4, pERK, ERK1, and ERK2 from Santa Cruz Biotechnology (Santa Cruz, CA, USA); ATF4 from GeneTex (Irvine, CA, USA).
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