Enhanced chemiluminescence system

Manufactured by Thermo Fisher Scientific

Sourced in United States, China, Germany, United Kingdom, Japan, Spain

The Enhanced chemiluminescence system is a laboratory instrument designed to detect and quantify specific proteins in biological samples. It utilizes a chemiluminescent detection method to produce a measurable light signal proportional to the amount of target protein present. The system provides a sensitive and reliable means for protein analysis in various research and diagnostic applications.

Automatically generated - may contain errors

Lab products found in correlation

Pvdf membrane, by Merck Group (24 mentions) Polyvinylidene fluoride membrane, by Merck Group (21 mentions) Polyvinylidene difluoride membrane, by Merck Group (12 mentions) Pvdf membrane, by Thermo Fisher Scientific (10 mentions) Protease inhibitor cocktail, by Roche (10 mentions) Protease inhibitor, by Roche (10 mentions) Ripa buffer, by Beyotime (9 mentions) Piercetm bca protein assay kit, by Thermo Fisher Scientific (9 mentions) Bca protein assay kit, by Thermo Fisher Scientific (9 mentions) Ripa buffer, by Thermo Fisher Scientific (8 mentions) β actin, by Santa Cruz Biotechnology (8 mentions) Ripa lysis buffer, by Beyotime (8 mentions) Bca protein assay kit, by Beyotime (8 mentions) Ripa buffer, by Merck Group (8 mentions) Quantity one software, by Bio-Rad (8 mentions) Pvdf membrane, by Bio-Rad (8 mentions) Bca kit, by Thermo Fisher Scientific (8 mentions) Anti β actin, by Cell Signaling Technology (7 mentions) Gapdh, by Cell Signaling Technology (7 mentions) Horseradish peroxidase conjugated secondary antibody, by Cell Signaling Technology (6 mentions)

285 protocols using enhanced chemiluminescence system

Quantitative Protein Analysis by Western Blot

Check if the same lab product or an alternative is used in the 5 most similar protocols

Radioimmunoprecipitation assay buffer (Beyotine, China) was used to extract protein; 15% sodium dodecyl sulfate-polyacrylamide gel electrophoresis was used for protein separation. Subsequently, we transferred proteins to polyvinylidene fluoride (PVDF) membrane. After blocking with BSA, the PVDF membrane was incubated with a specific antibody. The Enhanced Chemiluminescence System (Invitrogen, CA) was used to detect protein bands with specific antibodies. Anti-CDK1 (1:1000, Cat.ab18, Abcam, UK) and anti-β-actin (1:1000, Cat.ab8226, Abcam, UK) were primary antibodies. β-actin was considered as an endogenous control. Image J software was used to analyze the quantity of protein.

Zhu Y., Li K., Zhang J., Wang L., Sheng L, & Yan L. (2020). Inhibition of CDK1 Reverses the Resistance of 5-Fu in Colorectal Cancer. Cancer Management and Research, 12, 11271-11283.

+ Open protocol

+ Expand

Protein Expression Analysis in Lung Samples

Check if the same lab product or an alternative is used in the 5 most similar protocols

Lung samples and PMVECs were lysed in the radioimmunoprecipitation assay buffer containing 1% protease inhibitor (Beyotime Biotechnology, China). The cell lysates were centrifuged, and then supernatant was collected. The total protein concentration in the supernatant was quantified by BCA protein assay. Equal proteins were separated in 10% sodium dodecyl sulfate–polyacrylamide gel electrophoresis and then transferred to a polyvinylidene difluoride membrane. Primary antibodies specific for KLF6 (1:1000, Santa Cruz, sc-365633), ALK1 (1:1000, Abcam, ab108207), ENG (1:1000, R&D Systems, AF6440), PCNA (1:1000, Abcam, ab152112), GAPHD (1:1000, Abcam, ab8245), β-actin (1:1000, CST, #4970), and β-tubulin (1:1000, Abcam, ab6046) were diluted in 1% (w/v) bovine serum albumin in Tris-buffered saline with Tween (10 mM Tris-HCl, pH 7.5, 150 mM NaCl, 0.05% Tween-20), and the second antibody was diluted to 1:5000 (Abcam) in the same blocking buffer. The blots were visualized with enhanced chemiluminescence system (Invitrogen) and analyzed with GeneSnap (Syngene, Cambridge, United Kingdom).

Yang Y., Yu H., Yang C., Zhang Y., Ai X., Wang X., Lu K, & Yi B. (2019). Krüppel-like factor 6 mediates pulmonary angiogenesis in rat experimental hepatopulmonary syndrome and is aggravated by bone morphogenetic protein 9. Biology Open, 8(6), bio040121.

+ Open protocol

+ Expand

Protein Quantification and Western Blot Analysis

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Protein samples were gathered in a lysis buffer with protease inhibitors. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was utilized for protein separation. Then, the samples were transferred to polyvinylidene fluoride (PVDF) membranes incubated with a specific antibody [17 (link)]. The Enhanced Chemiluminescence System (Invitrogen, CA) was used to detect the Protein strip, which was semi-quantified using ImageJ (NIH, USA). This study included the primary antibodies of anti-TYRO3 (1:1000, Abcam, UK), anti-ENO1 (1:1000, Boster, China), anti-E-cadherin (1:1000, Bioss, China), anti-Vimentin (1:1000, Bioss, China) and anti-β-Actin (1:5000, Bioss, China). ImageJ software helped analyze the protein quantity.

Shao X., Sun Y., Zhong K., Gu J., Yu Y., Hu T., Kuai X, & Xing Y. (2023). TYRO3 promotes tumorigenesis and drug resistance in colorectal cancer by enhancing the epithelial-mesenchymal transition process. Aging (Albany NY), 15(8), 3035-3051.

+ Open protocol

+ Expand

Protein Expression Profiling in Prostate Cancer

Check if the same lab product or an alternative is used in the 5 most similar protocols

1 × 10⁶ PC-3 and E006AA prostate cancer cells were cultured for 24 hours, washed with cold PBS, and then lysed with SoluLyse-M (Genlantis, San Diego, CA) cell lysis Tris sucrose buffer. In addition to the cells, total protein was isolated from frozen prostate tumors and homogenized using the Millipore extraction kit (Millipore Corporation, Bilerica, MA). Proteins (30 and 50 μg) were separated using 10% or 16% pre-cast Tris-Glycine gels and dry-transferred for seven minutes using iBlot machine (Invitrogen, Gaithersburg, MD) onto PVDF membranes (Invitrogen, Gaithersburg, MD). The membrane was blocked using WesternBreeze Chemiluminescent Immunodetection Kit (Invitrogen) and probed with anti-Hsp27, Hsp40, Hsp60, Hsp70, Hsp90, NF-κB p65, VEGF, p53 (Ser15), p21 and cyclin D1 (1:500 diluted in manufacturer primary antibody diluent buffer) overnight at 4 °C. After washing with Invitrogen buffer wash (Invitrogen), the blots were treated with either Invitrogen Alk-Phos conjugated (anti-Mouse) or (anti-rabbit) for 30 minutes and washed several times. Proteins were detected by the enhanced chemiluminescence system (Invitrogen). Western analysis was performed on total cell lysate.

Ignacio D.N., Mason K.D., Hackett-Morton E.C., Albanese C., Ringer L., Wagner W.D., Wang P.C., Carducci M.A., Kachhap S.K., Paller C.J., Mendonca J., Li-Ying Chan L., Lin B., Hartle D.K., Green J.E., Brown C.A, & Hudson T.S. (2019). Muscadine grape skin extract inhibits prostate cancer cells by inducing cell-cycle arrest, and decreasing migration through heat shock protein 40. Heliyon, 5(1), e01128.

+ Open protocol

+ Expand

Exosome Protein Profiling Protocols

Check if the same lab product or an alternative is used in the 5 most similar protocols

Briefly, MSC and derived exosomes were lysed in lysis buffer, pH 7.4 (50 mM HEPES, 5 mM EDTA, and 50 mM NaCl), 1% Triton X-100, protease inhibitors (10 μg/mL aprotinin, 1 mM phenylmethylsulfonyl fluoride, and 10 μg/mL leupeptin), and phosphatase inhibitors (50 mM sodium fluoride, 1 mM sodium orthovanadate, and 10 mM sodium pyrophosphate). The protein samples (40 μg) were electrophoresed using SDS-polyacrylamide gel and electroimmunoblotted as described [8 (link)]. After protein transfer, the samples on PVDF membranes were visualized using an enhanced chemiluminescence system (Invitrogen), exposed to X-ray film, and then quantified by laser scanning densitometry. The specific antibodies used for detection of antigens of interest are listed in the following: anti-CXCR4 (Millipore), anti-Akt, anti-pAkt (Cell Signal Technology), anti-IGF-1α, anti-CD9, anti-CD63, and anti-β-actin antibody (Santa Cruz Biotechnology).

Kang K., Ma R., Cai W., Huang W., Paul C., Liang J., Wang Y., Zhao T., Kim H.W., Xu M., Millard R.W., Wen Z, & Wang Y. (2015). Exosomes Secreted from CXCR4 Overexpressing Mesenchymal Stem Cells Promote Cardioprotection via Akt Signaling Pathway following Myocardial Infarction. Stem Cells International, 2015, 659890.

+ Open protocol

+ Expand

Western Blot Protein Expression Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells were lysed in M-PER Mammalian Protein Extraction Reagent (Thermo SCIENTIFIC, Rockford, IL, USA), 1X protease inhibitor cocktail (Calbiochem, Darmstadt, Germany) and 1X phosphatase inhibitor cocktail 2 & 3 (Sigma-Aldrich, St. Louis, MO, USA), sonicated for 10 seconds, followed by centrifugation (13,200 rpm, 4C, 5 min), after which the supernatants were stored at −80C until use. Protein concentrations were determined with the Pierce^® BCA Protein Assay kit (Thermo SCIENTIFIC), using BSA as the concentration standard. Extracted proteins were then resolved using 4–12% Bis-Tris Gels (Life Technologies, Carlsbad, CA, USA) and subsequently transferred to PVDF membranes (Life Technologies). Membranes were washed and blocked with 1% BSA in TBS (20mM Tris-HCl, 150mM NaCl, pH 7.4) containing 0.1% Tween 20 (TBS-T) before incubation with primary antibodies. Next, they were incubated with horseradish peroxidase-conjugated donkey anti-rabbit IgG or sheep anti-mouse IgG (GE Healthcare). Finally, the signals were visualized by means of an enhanced chemiluminescence system (Invitrogen, Carlsbad, CA, USA). The primary antibodies were used at the following dilutions: anti-phospho-p44/42MAPK (1:1,000), anti-p44/42MAPK (1:1,000), and anti-GAPDH (1:2,000) (Cell Signaling Technology, Danvers, MA) antibodies. anti-GAPDH antibody was used as a loading control.

Blehm B.H., Jiang N., Kotobuki Y, & Tanner K. (2015). Deconstructing the role of the ECM microenvironment on drug efficacy targeting MAPK signaling in a pre-clinical platform for cutaneous melanoma. Biomaterials, 56, 129-139.

+ Open protocol

+ Expand

Protein Expression Analysis of Mammary Epithelial Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Total protein was isolated from frozen, Balb/C, SP3 and NSP2, mammary epithelial cells and homogenized using the Millipore extraction kit (Millipore Corporation, Billerica, MA). Proteins (30 and 50μg) were separated using 10% or 16% pre-cast Tris-Glycine gels and dry-transferred for seven minutes using the iBlot machine (Invitrogen, Gaithersburg, MD) onto PVDF membranes (Invitrogen, Gaithersburg, MD). The membrane was blocked using WesternBreeze Chemiluminescent Immunodetection Kit (Invitrogen) and probed with anti-ID2 (Abcam-ab85990), SOX9 (Abcam-ab3697), cytokeratin 5 (Abcam-ab53121), and cytokeratin 8 (Abcam-ab154301) (1:500 diluted in manufacturer primary antibody diluent buffer) overnight at 4°C. The additional probing with beta-actin (Chemicon) antibody used as an internal control. After washing with Invitrogen buffer wash (Invitrogen), the blots were treated with either Invitrogen Alk-Phos conjugated (anti-Mouse) or (anti-rabbit) for 30 minutes and washed several times. Proteins were detected by the enhanced chemiluminescence system (Invitrogen). Western analysis was performed on total cell lysate.

Johnson J.R., Boulanger C.A., Hudson T., Savage E, & Smith G.H. (2019). Microarray and pathway analysis of two COMMA-Dβ derived clones reveal important differences relevant to their developmental capacity in-vivo. Oncotarget, 10(22), 2118-2135.

+ Open protocol

+ Expand

Western Blot Analysis of Astrocyte Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols

Astrocytes were lysed in RIPA buffer (Thermo Fisher Scientific K.K. Tokyo, Japan, 25 mM Tris-HCl pH 7.6, 150 mM NaCl, 1% NP-40, 1% sodium deoxycholate, and 0.1% SDS) containing a complete protease inhibitor cocktail (Roche Diagnostics). Cell lysates (30 μg protein/line) were assessed by SDS-PAGE and transferred to polyvinylidene difluoride membranes. Membranes were treated with a horseradish peroxidase-conjugated antibody (Life Technologies), and expression levels were examined using an enhanced chemiluminescence system (Life Technologies) with a C-Digit blot scanner (MS Techno Systems Inc. Tokyo, Japan). Antibodies for anti-MCT1 antibody (1: 200, Santa Cruz Biotechnology, USA: sc-50325), anti-MCT4 (1:200, Santa Cruz Biotechnology, USA: sc-50329), anti-iNOS (1:500, Abcam, ab 3523), anti-eNOS (1:1000, Cell Signaling Technology, The Netherlands: #32027), and β-actin (1:1000, Cell Signaling Technology: #4967) were used.

Yamagata K., Takahashi N., Akita N, & Nabika T. (2017). Arginine vasopressin altered the expression of monocarboxylate transporters in cultured astrocytes isolated from stroke-prone spontaneously hypertensive rats and congenic SHRpch1_18 rats. Journal of Neuroinflammation, 14, 176.

+ Open protocol

+ Expand

Western Blot Analysis of Molecular Targets

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Whole protein extracts and Western blot (WB) analysis were performed using standard procedures as described in our previous study,^{9 (link)} with detection using the enhanced chemiluminescence system (Life Technologies, Grand Island, NY). Antibodies against SLC1A5 were used at 1:1,000 dilutions (Millipore, Billerica, MA), SN1 at 1:100 (Epitomics, Burlin-game, CA), LAT1 at 1:1,000 (Cell Signaling Technology, Danvers, MA), Caspase 3 at 1:1,000 (Cell Signaling Technology), Caspase 9 at 1:1,000 (Cell Signaling Technology), LC3 1:1,000 (Medical and Biological Laboratories, Nagoya, Japan), Bcl-2 1:1,000 (Cell Signaling Technology) and Bax 1:1000 (Cell Signaling Technology).

Hassanein M., Qian J., Hoeksema M.D., Wang J., Jacobovitz M., Ji X., Harris F.T., Harris B.K., Boyd K.L., Chen H., Eisenberg R, & Massion P.P. (2015). Targeting SLC1A5-mediated glutamine dependence in non-small cell lung cancer. International journal of cancer. Journal international du cancer, 137(7), 1587-1597.

+ Open protocol

+ Expand

Western Blot Analysis of Astrocyte Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols

Astrocytes were lysed in RIPA buffer and cell lysates (15-30 μg protein/line) were separated by SDS-PAGE and transferred to polyvinylidene difluoride membranes, as described previously (Yamagata et al. 2013) . Membranes were then incubated with primary antibodies (anti-β-actin, 1:2000, Cell Signaling Technology, Tokyo, Japan; or anti-Hmgb1, 1:10,000, Abcam, EPR3507) for 1.5 h or 5 h respectively. After being washed twice, membranes were incubated with a horseradish peroxidase-conjugated secondary antibody (Life Technologies), and protein levels were detected using an enhanced chemiluminescence system (Life Technologies) with ChemiDoc XRS Plus (Bio-Rad, Tokyo, Japan).

Yamagata K., Sone N., Suguyama S, & Nabika T. (2016). Different effects of arginine vasopressin on high-mobility group box 1 expression in astrocytes isolated from stroke-prone spontaneously hypertensive rats and congenic SHRpch1_18 rats. International journal of experimental pathology, 97(2).

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!