Enhanced chemiluminescence ecl kit

Manufactured by PerkinElmer

Sourced in United States

The Enhanced chemiluminescence (ECL) kit is a laboratory product that detects and quantifies specific proteins in a sample. It uses a chemiluminescent reaction to generate a measurable light signal proportional to the amount of target protein present.

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Lab products found in correlation

Dimethyl sulfoxide (dmso), by Merck Group (2 mentions) Mitochondrial membrane potential detection kit, by Beyotime (1 mentions) D luciferin potassium salt, by Yeasen (1 mentions) Annexin 5 fitc pi apoptosis detection kit, by Beyotime (1 mentions) Cell counting kit 8 cck 8 assay kit, by MedChemExpress (1 mentions) Cdk7 inhibitor thz1, by MedChemExpress (1 mentions) Cell culture medium, by Thermo Fisher Scientific (1 mentions) Fluo 4 am, by Yeasen (1 mentions) Trypsin edta, by Thermo Fisher Scientific (1 mentions) P akt, by Affinity Biosciences (1 mentions) Annexin 5 fitc pi apoptosis detection kit, by BD (1 mentions) Cell counting kit 8 cck 8, by BD (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions) Penicillin streptomycin, by Merck Group (1 mentions) Acetonitrile, by Merck Group (1 mentions) Hplc grade methanol, by Merck Group (1 mentions) Bcl 2, by Abcam (1 mentions) Caspase 3, by Abcam (1 mentions) Beas 2b, by National Collection of Authenticated Cell Cultures (1 mentions)

5 protocols using enhanced chemiluminescence ecl kit

Characterization of Apoptosis Induction

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Calcium chloride dihydrate (CaCl₂·2H₂O), alendronate sodium trihydrate (NaALN·3H₂O) were purchased from Shanghai Aladdin Bio-Chem Technology Co., LTD (Shanghai, China). CDK7 inhibitor THZ1 and the cell counting kit-8 (CCK8) assay kit were purchased from MedChemExpress (MCE, Shanghai). Dimethyl sulfoxide (DMSO) and penicillin–streptomycin (pen/strep) solution (100 ×) were purchased from Sigma Aldrich (MO, USA). Cell culture medium, trypsin–EDTA and fetal bovine serum (FBS) were provided by Gibco (Guangzhou, China). Annexin V-FITC/PI apoptosis detection kits and mitochondrial membrane potential detection kit were purchased from Beyotime (Shanghai, China). 6-carboxy fluorescein (6-FAM), DAPI dihydrochloride and DiR iodide were purchased from Invitrogen (Carlsbad, CA, USA). D-Luciferin potassium salt and Fluo-4, AM were acquired from Yeasen Biotech Co., Ltd (Shanghai, China). Acridine orange-ethidium bromide (AO/EB) staining kit was purchased from Leagene Biotechnology (Beijing, China). Bcl-2, Bax and cleaved caspase-3 antibodies were bought from Absin Bioscience (Shanghai, China). β-actin and secondary antibodies were purchased from Proteintech (Wuhan, China). BCA protein quantitation kit and enhanced chemiluminescence (ECL) kit were purchased from Perkin-Elmer (Waltham, USA). All other reagents were consistent with previous report [35 (link)].

Huang X., Qiu M., Wang T., Li B., Zhang S., Zhang T., Liu P., Wang Q., Qian Z.R., Zhu C., Wu M, & Zhao J. (2022). Carrier-free multifunctional nanomedicine for intraperitoneal disseminated ovarian cancer therapy. Journal of Nanobiotechnology, 20, 93.

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Anticancer Properties of C. tinctoria

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The C. tinctoria was from the research base of the Uyghur Autonomous Region Institute of Medicinal Plants (Xinjiang, China). Human lung carcinoma (A549 and NCI-H292) cells and BEAS-2B were purchased from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China). RPMI-1640 medium and fetal bovine serum (FBS) was from Gibco (United States). Penicillin/streptomycin, and dimethyl sulfoxide (DMSO) were purchased from Sigma (Missouri, United States). Cell Counting Kit-8 (CCK-8) and Annexin V-FITC/PI apoptosis detection kits were from BD Biosciences (United States). HPLC grade methanol and acetonitrile were purchased from Merck (Darmstadt, Germany). The antibodies of Caspase-3, Bax, and Bcl-2 were all obtained from Abcam (United States); those of p-PI3K, PI3K, p-Akt, and Akt were from Affinity Biosciences (Cincinnati, United States); and, that of β-actin was from Proteintech Group (Wuhan, China). An enhanced chemiluminescence (ECL) kit was purchased from PerkinElmer (Waltham, United States).

Wufuer Y., Yang X., Guo L., Aximujiang K., Zhong L., Yunusi K, & Wu G. (2022). The Antitumor Effect and Mechanism of Total Flavonoids From Coreopsis Tinctoria Nutt (Snow Chrysanthemum) on Lung Cancer Using Network Pharmacology and Molecular Docking. Frontiers in Pharmacology, 13, 761785.

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Western Blot Analysis of CEA, TGFβ Signaling

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Cells were lysed with lysis buffer and were standardized using a BCA protein assay (Pierce™ BCA Protein Assay Kit 23225). Equal amounts of proteins were fractionated on SDS–PAGE and blotted to nitrocellulose membrane. Membranes were incubated with CEA (Thermo MS-613-P0), TGFBR1 (Santa Cruz Sc-398), TGFBR2 (ab17650), Phospho-Smad3 (Ser423/425) (Cell Signaling #9520), Smad3 (Cell Signaling #9523), and tubulin (Cell Signaling #2144) antibodies. Secondary antibodies conjugated with horseradish peroxidase (Chemicon) and Enhanced chemiluminescence (ECL) kit (Perkin-Elmer Life Sciences) were used to develop the immunoblots.

Chen J., Raju G.S., Jogunoori W., Menon V., Majumdar A., Chen J.S., Gi Y.J., Jeong Y.S., Phan L., Belkin M., Gu S., Kundra S., Mistry N.A., Zhang J., Su X., Li S., Lin S.H., Javle M., McMurray J.S., Rahlfs T.F., Mishra B., White J., Rashid A., Beauchemin N., Weston B.R., Shafi M.A., Stroehlein J.R., Davila M., Akbani R., Weinstein J.N., Wu X, & Mishra L. (2016). Mutational Profiles Reveal an Aberrant TGF-β-CEA Regulated Pathway in Colon Adenomas. PLoS ONE, 11(4), e0153933.

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PBrP Modulation of TGF-β Signaling

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To examine the effect of PBrP on TGF-β-induced Smad2/3 phosphorylation, Mv1Lu and A549 cells were incubated with PBrP at various concentrations or for various periods and then stimulated with 100 pM of TGF-β for 30 min. To monitor EMT protein expression, the cells were pretreated with different PBrP concentrations for 2 h and then stimulated with TGF-β for 48 h. The cells were lysed with a lysis buffer and standardised using a bicinchoninic acid (BCA) protein assay (Pierce™ BCA Protein Assay Kit 23225). Fifty microgram of this protein was applied to SDS-PAGE and electrotransferred to the polyvinylidene difluoride (PVDF) membrane. Polyclonal antibodies against TβRI (sc-398), TβRII (sc-400), EGFR (sc-373746), β-actin (sc-47778), flotilin-1 (sc-25506), fibronectin (sc-9068), N-cadherin (sc-7939), Lamin B (sc-6216) and vimentin (sc-7557) were obtained from Santa Cruz (Dallas, TX). Rabbit Polyclonal antibodies against pSmad2/3 (#8828), Smad2/3 (#8685), PAI-1 (#11907), Flag tag (#8146), Lamp-1 (sc-9091) and MyoVa (#3402) were purchased from Cell Signalling (Boston, MA). Secondary antibodies conjugated with horseradish peroxidase (Millipore, Darmstadt, Germany) and an enhanced chemiluminescence (ECL) kit (Perkin-Elmer Life Sciences, Waltham, MA, USA) were used to develop immunoblots.

Shih-Wei W., Chih-Ling C., Kao Y.C., Martin R., Knölker H.J., Shiao M.S, & Chen C.L. (2018). Pentabromopseudilin: a myosin V inhibitor suppresses TGF-β activity by recruiting the type II TGF-β receptor to lysosomal degradation. Journal of Enzyme Inhibition and Medicinal Chemistry, 33(1), 920-935.

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Lipid Raft Disruption and Antibody Analysis

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Methyl-β-cyclodextr in (MβCD) and anti-neutrophil myeloperoxidase [MPO; mouse anti-goat polyclonal IgG (H+L)] antibodies were obtained from Santa Cruz Biotechnology, Inc. (1:100, cat. no. sc-16129, Dallas, TX, USA), along with lipid-raft disruptor filipin. Alexa Fluor 488-cholera toxin subunit B (CTXB) and Alexa Fluor 594-labeled chicken anti-Goat IgG (H+L) secondary antibody were purchased from Invitrogen (Thermo Fisher Scientific, Inc., Waltham, MA, USA). Anti-flotillin-1 antibody was purchased from BD Biosciences (1:1000, cat no. 61802; Franklin Lakes, NJ, USA). Polyclonal anti-ERp29 rabbit anti-mouse antibody was obtained from Abcam (1:3000, ab11420; Cambridge, MA, USA). Optiprep was obtained from Axis-Shield, Inc. (Norton, MA, USA). High glucose Dulbecco's modified Eagle's medium (DMEM) was purchased from GE Healthcare Life Sciences (Logan, UT, USA). An Enhanced Chemiluminescence (ECL) kit was obtained from PerkinElmer Inc. (Waltham, MA, USA). Human LDL was purchased from ProSpec-Tany TechnoGene, Ltd. (Ness Ziona, Israel).

SONG J., PING L.Y., DUONG D.M., GAO X.Y., HE C.Y., WEI L, & WU J.Z. (2016). Native low density lipoprotein promotes lipid raft formation in macrophages. Molecular Medicine Reports, 13(3), 2087-2093.

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