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Amresco cycloheximide 94271
Cycloheximide 94271, supplied by Amresco, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/94271/cycloheximide+94271/pm38701646-357-12-14
Average 90 stars, based on 1 article reviews
cycloheximide 94271 - by Bioz Stars, 2026-11
90/100 stars

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Related Articles

Transfection:

Article Title: CERKL regulates autophagy via the NAD-dependent deacetylase SIRT1
Article Snippet: After 72 h, cell lysates were harvested and immunoprecipitated with pan acetyl-lysine antibody (ABclonal, A2391) and protein G beads, and washed and analyzed by western blotting with the antibody. .. For protein half-life experiments cells were treated with 200 μg/ml or 300 μg/ml cycloheximide (Amresco, 94271) 12 h following transfection with siRNAs or plasmids and subsequently sampled at the indicated times. ..

Incubation:

Article Title: The absence of the ribosomal protein Rpl2702 elicits the MAPK-mTOR signaling to modulate mitochondrial morphology and functions.
Article Snippet: The samples were separated by SDS-PAGE and then analyzed by immunoblotting with antibodies against GFP (dilution factor, 1:2000; 600-101-215, Rockland), Mti3 (dilution factor, 1:2000; Homemade) and Tubulin (dilution factor, 1:10,000; 63–160, Bio Academia). .. Cells were incubated in EMM5S overnight until OD600 reached 0.6–0.8, 100 μg/mL cycloheximide (94271, Amresco) was added to the medium for 15 min at 30 ◦C, and 100 mL cells were collected and washed three times with Diethyl pyrocarbonate (DEPC)-treated water (B600154-0025, Sangon). .. Subsequently, the cells were resuspended in ice-cold 600 μL lysis buffer (20 mM Tris-HCl (pH 7.5), 150 mM NaCl, 5 mM MgCl2, 1 % Triton X-100, 200 μg/mL cycloheximide, 20 units/mL RNase inhibitors, cocktail protease inhibitors (EDTA-free), and 1 mM dithiothreitol), and lysed with 200 μL ice-cold 0.5 mm glass beads (BioSpec) using a mixer mill (Retsch MM400) (www.retsch.com).

Inhibition:

Article Title: CCDC84 Acetylation Oscillation Regulates Centrosome Duplication by Modulating HsSAS-6 Degradation.
Article Snippet: For the S-phase arrest-induced centriole overduplication assays, 2 mM hydroxyurea (#H8627, Sigma-Aldrich) was added to U2OS cells for 36 h, or 1.5 mg/mL aphidicolin (#178273, Millipore) was added to HeLa cells for 36 h. For inhibition of HDAC or SIRT1 activity, HeLa or U2OS cells were treated with 100 ng/mL trichostatin (#T1952, Sigma-Aldrich) or 1 mM Ex-527 (#S1541, Selleckchem), respectively, for 48 h. For inhibition of NAT10 activity, HeLa cells were treated with 20 mM remodelin (#S7641, Selleckchem) for 48 h. For inhibition of proteasome activity, HeLa cells were treated with 50 mg/mL MG132 (#M8699, Sigma-Aldrich) for 6 h. For measurement of the HsSAS-6 half-life, HeLa cells were treated with 50 mg/mL cycloheximide (#94271, Amresco). .. For the S-phase arrest-induced centriole overduplication assays, 2 mM hydroxyurea (#H8627, Sigma-Aldrich) was added to U2OS cells for 36 h, or 1.5 mg/mL aphidicolin (#178273, Millipore) was added to HeLa cells for 36 h. For inhibition of HDAC or SIRT1 activity, HeLa or U2OS cells were treated with 100 ng/mL trichostatin (#T1952, Sigma-Aldrich) or 1 mM Ex-527 (#S1541, Selleckchem), respectively, for 48 h. For inhibition of NAT10 activity, HeLa cells were treated with 20 mM remodelin (#S7641, Selleckchem) for 48 h. For inhibition of proteasome activity, HeLa cells were treated with 50 mg/mL MG132 (#M8699, Sigma-Aldrich) for 6 h. For measurement of the HsSAS-6 half-life, HeLa cells were treated with 50 mg/mL cycloheximide (#94271, Amresco). ..

Activity Assay:

Article Title: CCDC84 Acetylation Oscillation Regulates Centrosome Duplication by Modulating HsSAS-6 Degradation.
Article Snippet: For the S-phase arrest-induced centriole overduplication assays, 2 mM hydroxyurea (#H8627, Sigma-Aldrich) was added to U2OS cells for 36 h, or 1.5 mg/mL aphidicolin (#178273, Millipore) was added to HeLa cells for 36 h. For inhibition of HDAC or SIRT1 activity, HeLa or U2OS cells were treated with 100 ng/mL trichostatin (#T1952, Sigma-Aldrich) or 1 mM Ex-527 (#S1541, Selleckchem), respectively, for 48 h. For inhibition of NAT10 activity, HeLa cells were treated with 20 mM remodelin (#S7641, Selleckchem) for 48 h. For inhibition of proteasome activity, HeLa cells were treated with 50 mg/mL MG132 (#M8699, Sigma-Aldrich) for 6 h. For measurement of the HsSAS-6 half-life, HeLa cells were treated with 50 mg/mL cycloheximide (#94271, Amresco). .. For the S-phase arrest-induced centriole overduplication assays, 2 mM hydroxyurea (#H8627, Sigma-Aldrich) was added to U2OS cells for 36 h, or 1.5 mg/mL aphidicolin (#178273, Millipore) was added to HeLa cells for 36 h. For inhibition of HDAC or SIRT1 activity, HeLa or U2OS cells were treated with 100 ng/mL trichostatin (#T1952, Sigma-Aldrich) or 1 mM Ex-527 (#S1541, Selleckchem), respectively, for 48 h. For inhibition of NAT10 activity, HeLa cells were treated with 20 mM remodelin (#S7641, Selleckchem) for 48 h. For inhibition of proteasome activity, HeLa cells were treated with 50 mg/mL MG132 (#M8699, Sigma-Aldrich) for 6 h. For measurement of the HsSAS-6 half-life, HeLa cells were treated with 50 mg/mL cycloheximide (#94271, Amresco). ..

other:

Article Title: Accurate, Streamlined Analysis of mRNA Translation by Sucrose Gradient Fractionation
Article Snippet: Triton X-100 solution (Sigma-Aldrich, catalog number: 93443) Dithiothreitol (DTT) (Sigma-Aldrich, catalog number: D9779) Note: Store filtered and aliquotted 1 M stock solution at -20 °C.

Article Title: Use of Antibiotics for Maintenance of Axenic Cultures of Amphidinium carterae for the Analysis of Translation
Article Snippet: Cycloheximide (94271) was from Amresco (Solon, OH, USA).

Article Title: Leucine Protects Against Skeletal Muscle Atrophy in Lipopolysaccharide-Challenged Rats.
Article Snippet: Skeletal muscle atrophy is a decrease in muscle mass that occurs when protein degradation exceeds protein synthesis.. Leucine (Leu), an essential branched-chain amino acid in animal nutrition, regulates skeletal muscle protein metabolism.. Two experiments were conducted to evaluate whether Leu could alleviate lipopolysaccharide (LPS)-induced skeletal muscle wasting by modulating skeletal muscle protein synthesis and degradation.



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CDC34 inhibits the ubiquitination of EGFR and protects it from proteolytic degradation. (a) qRT-PCR analysis of CDC34 and EGFR in A549 and H1975 cells 72 h after si CDC34 transfection. Error bars, sd; P values, Student's t -test. (b) <t>Cycloheximide</t> <t>(CHX)</t> chase assay to measure EGFR half-life in H460 (in the absence and presence of EGF) and H1975 cells transfected with si CDC34- 1# (lower panel). The relative expression values of EGFR were the results determined by densitometry analysis normalized to Actin. Error bars, sd. (c) HCC827 cells were transfected with 50 nM si CDC34- 1# for 24 h, treated with erlotinib for 24 h, and lyszed for Western blot assays using indicated antibodies. (d) Western blot analysis of the EGFR expression in H460 cells transfected with si CDC34- 1# in the presence or absence of MG132. (e) A549 and H1975 cells were transfected with si CDC34 and HA-Ub, lysed, and the lysates were subjected to immunoprecipitation (IP) and immunoblot using indicated antibodies. (f) A549 cells were transfected with Flag- CDC34 and HA-Ub, and lysed for IP and immunoblot assays. (g) H460 cells were transfected with indicated constructs, treated with MG132, and lysed for IP and immunoblot assays. (h) The cells were transfected with CDC34 and HA-Ub, and lysed for IP and immunoblot assays using indicated antibodies. (i) Western blot analysis of EGFR in #1 si CDC34 -transfected H460 cells in the presence or absence of chloroquine (CQ). (j) HCC827 cells were transfected with siCDC34 and HA-Ub for 24 h, treated with or without chloropuine for additional 24 h, lyzed, and subjected to IP and immunoblot using indicated antibodies. (k) H460, A549, and H1975 cells transfected with si CDC34 were lysed and subjected to Co-IP and immunoblotting using indicated antibodies. (l) H460 cells were transfected with CDC34 (left) or CBL (right) in the absence or presence of EGF for 48 h, and lysed for Co-IP and immunoblot using indicated antibodies. (m) In vitro Co-IP experiments using Flag-EGFR purified from 293T cells, His-CDC34 (from E. coli), and increasing amount of purified His-c-Cbl (from E. coli) proteins and indicated antibodies. (n) In vitro Co-IP experiments using Flag-EGFR purified from 293T cells, His-c-Cbl (from E. coli), and increasing amount of purified His-CDC34 (from E. coli) proteins and indicated antibodies. (o) 293T cells were transfected with the wild type or mutant (Y1045F) Flag- EGFR ICD for 48 h, lyzed, and subjected to IP and immunobloting using indicated antibodies. (p) si CDC34 -mediated loss of EGFR is rescued by knockdown of CBL . A549 cells were transfected with si CDC34 and/or si CBL for 72 h. Before lysis for immunoblot assays, the cells were serum-starved for 3 h and then stimulated by 50 ng/mL EGF for 15 min.
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CDC34 inhibits the ubiquitination of EGFR and protects it from proteolytic degradation. (a) qRT-PCR analysis of CDC34 and EGFR in A549 and H1975 cells 72 h after si CDC34 transfection. Error bars, sd; P values, Student's t -test. (b) <t>Cycloheximide</t> <t>(CHX)</t> chase assay to measure EGFR half-life in H460 (in the absence and presence of EGF) and H1975 cells transfected with si CDC34- 1# (lower panel). The relative expression values of EGFR were the results determined by densitometry analysis normalized to Actin. Error bars, sd. (c) HCC827 cells were transfected with 50 nM si CDC34- 1# for 24 h, treated with erlotinib for 24 h, and lyszed for Western blot assays using indicated antibodies. (d) Western blot analysis of the EGFR expression in H460 cells transfected with si CDC34- 1# in the presence or absence of MG132. (e) A549 and H1975 cells were transfected with si CDC34 and HA-Ub, lysed, and the lysates were subjected to immunoprecipitation (IP) and immunoblot using indicated antibodies. (f) A549 cells were transfected with Flag- CDC34 and HA-Ub, and lysed for IP and immunoblot assays. (g) H460 cells were transfected with indicated constructs, treated with MG132, and lysed for IP and immunoblot assays. (h) The cells were transfected with CDC34 and HA-Ub, and lysed for IP and immunoblot assays using indicated antibodies. (i) Western blot analysis of EGFR in #1 si CDC34 -transfected H460 cells in the presence or absence of chloroquine (CQ). (j) HCC827 cells were transfected with siCDC34 and HA-Ub for 24 h, treated with or without chloropuine for additional 24 h, lyzed, and subjected to IP and immunoblot using indicated antibodies. (k) H460, A549, and H1975 cells transfected with si CDC34 were lysed and subjected to Co-IP and immunoblotting using indicated antibodies. (l) H460 cells were transfected with CDC34 (left) or CBL (right) in the absence or presence of EGF for 48 h, and lysed for Co-IP and immunoblot using indicated antibodies. (m) In vitro Co-IP experiments using Flag-EGFR purified from 293T cells, His-CDC34 (from E. coli), and increasing amount of purified His-c-Cbl (from E. coli) proteins and indicated antibodies. (n) In vitro Co-IP experiments using Flag-EGFR purified from 293T cells, His-c-Cbl (from E. coli), and increasing amount of purified His-CDC34 (from E. coli) proteins and indicated antibodies. (o) 293T cells were transfected with the wild type or mutant (Y1045F) Flag- EGFR ICD for 48 h, lyzed, and subjected to IP and immunobloting using indicated antibodies. (p) si CDC34 -mediated loss of EGFR is rescued by knockdown of CBL . A549 cells were transfected with si CDC34 and/or si CBL for 72 h. Before lysis for immunoblot assays, the cells were serum-starved for 3 h and then stimulated by 50 ng/mL EGF for 15 min.
Cycloheximide Amresco 94271, supplied by Amresco, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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CDC34 inhibits the ubiquitination of EGFR and protects it from proteolytic degradation. (a) qRT-PCR analysis of CDC34 and EGFR in A549 and H1975 cells 72 h after si CDC34 transfection. Error bars, sd; P values, Student's t -test. (b) Cycloheximide (CHX) chase assay to measure EGFR half-life in H460 (in the absence and presence of EGF) and H1975 cells transfected with si CDC34- 1# (lower panel). The relative expression values of EGFR were the results determined by densitometry analysis normalized to Actin. Error bars, sd. (c) HCC827 cells were transfected with 50 nM si CDC34- 1# for 24 h, treated with erlotinib for 24 h, and lyszed for Western blot assays using indicated antibodies. (d) Western blot analysis of the EGFR expression in H460 cells transfected with si CDC34- 1# in the presence or absence of MG132. (e) A549 and H1975 cells were transfected with si CDC34 and HA-Ub, lysed, and the lysates were subjected to immunoprecipitation (IP) and immunoblot using indicated antibodies. (f) A549 cells were transfected with Flag- CDC34 and HA-Ub, and lysed for IP and immunoblot assays. (g) H460 cells were transfected with indicated constructs, treated with MG132, and lysed for IP and immunoblot assays. (h) The cells were transfected with CDC34 and HA-Ub, and lysed for IP and immunoblot assays using indicated antibodies. (i) Western blot analysis of EGFR in #1 si CDC34 -transfected H460 cells in the presence or absence of chloroquine (CQ). (j) HCC827 cells were transfected with siCDC34 and HA-Ub for 24 h, treated with or without chloropuine for additional 24 h, lyzed, and subjected to IP and immunoblot using indicated antibodies. (k) H460, A549, and H1975 cells transfected with si CDC34 were lysed and subjected to Co-IP and immunoblotting using indicated antibodies. (l) H460 cells were transfected with CDC34 (left) or CBL (right) in the absence or presence of EGF for 48 h, and lysed for Co-IP and immunoblot using indicated antibodies. (m) In vitro Co-IP experiments using Flag-EGFR purified from 293T cells, His-CDC34 (from E. coli), and increasing amount of purified His-c-Cbl (from E. coli) proteins and indicated antibodies. (n) In vitro Co-IP experiments using Flag-EGFR purified from 293T cells, His-c-Cbl (from E. coli), and increasing amount of purified His-CDC34 (from E. coli) proteins and indicated antibodies. (o) 293T cells were transfected with the wild type or mutant (Y1045F) Flag- EGFR ICD for 48 h, lyzed, and subjected to IP and immunobloting using indicated antibodies. (p) si CDC34 -mediated loss of EGFR is rescued by knockdown of CBL . A549 cells were transfected with si CDC34 and/or si CBL for 72 h. Before lysis for immunoblot assays, the cells were serum-starved for 3 h and then stimulated by 50 ng/mL EGF for 15 min.

Journal: EBioMedicine

Article Title: Systematic identification of CDC34 that functions to stabilize EGFR and promote lung carcinogenesis

doi: 10.1016/j.ebiom.2020.102689

Figure Lengend Snippet: CDC34 inhibits the ubiquitination of EGFR and protects it from proteolytic degradation. (a) qRT-PCR analysis of CDC34 and EGFR in A549 and H1975 cells 72 h after si CDC34 transfection. Error bars, sd; P values, Student's t -test. (b) Cycloheximide (CHX) chase assay to measure EGFR half-life in H460 (in the absence and presence of EGF) and H1975 cells transfected with si CDC34- 1# (lower panel). The relative expression values of EGFR were the results determined by densitometry analysis normalized to Actin. Error bars, sd. (c) HCC827 cells were transfected with 50 nM si CDC34- 1# for 24 h, treated with erlotinib for 24 h, and lyszed for Western blot assays using indicated antibodies. (d) Western blot analysis of the EGFR expression in H460 cells transfected with si CDC34- 1# in the presence or absence of MG132. (e) A549 and H1975 cells were transfected with si CDC34 and HA-Ub, lysed, and the lysates were subjected to immunoprecipitation (IP) and immunoblot using indicated antibodies. (f) A549 cells were transfected with Flag- CDC34 and HA-Ub, and lysed for IP and immunoblot assays. (g) H460 cells were transfected with indicated constructs, treated with MG132, and lysed for IP and immunoblot assays. (h) The cells were transfected with CDC34 and HA-Ub, and lysed for IP and immunoblot assays using indicated antibodies. (i) Western blot analysis of EGFR in #1 si CDC34 -transfected H460 cells in the presence or absence of chloroquine (CQ). (j) HCC827 cells were transfected with siCDC34 and HA-Ub for 24 h, treated with or without chloropuine for additional 24 h, lyzed, and subjected to IP and immunoblot using indicated antibodies. (k) H460, A549, and H1975 cells transfected with si CDC34 were lysed and subjected to Co-IP and immunoblotting using indicated antibodies. (l) H460 cells were transfected with CDC34 (left) or CBL (right) in the absence or presence of EGF for 48 h, and lysed for Co-IP and immunoblot using indicated antibodies. (m) In vitro Co-IP experiments using Flag-EGFR purified from 293T cells, His-CDC34 (from E. coli), and increasing amount of purified His-c-Cbl (from E. coli) proteins and indicated antibodies. (n) In vitro Co-IP experiments using Flag-EGFR purified from 293T cells, His-c-Cbl (from E. coli), and increasing amount of purified His-CDC34 (from E. coli) proteins and indicated antibodies. (o) 293T cells were transfected with the wild type or mutant (Y1045F) Flag- EGFR ICD for 48 h, lyzed, and subjected to IP and immunobloting using indicated antibodies. (p) si CDC34 -mediated loss of EGFR is rescued by knockdown of CBL . A549 cells were transfected with si CDC34 and/or si CBL for 72 h. Before lysis for immunoblot assays, the cells were serum-starved for 3 h and then stimulated by 50 ng/mL EGF for 15 min.

Article Snippet: Reagents used included cycloheximide (CHX) (#94271, Amresco Inc., Solon, OH, USA), erlotinib (#HY-12008, MedChemExpress, USA), epoxomicin (#A2606, APExBIO, USA), Universal Tyrosine Kinase Assay Kit (#MK410, Clontech, Palo Alto, CA), MG132 (Sigma, #SML1135), chloroquine (Sigma, #PHR-1258), BaP (Sigma, #B1760), BAA (Sigma, #B2209), and DBA (Sigma, #91861).

Techniques: Ubiquitin Proteomics, Quantitative RT-PCR, Transfection, Expressing, Western Blot, Immunoprecipitation, Construct, Co-Immunoprecipitation Assay, In Vitro, Purification, Mutagenesis, Knockdown, Lysis