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Ccd camera based imager

Manufactured by GE Healthcare
Sourced in United States

The CCD (Charge-Coupled Device) camera-based imager is a lab equipment product designed for image capture and analysis. It utilizes a CCD sensor to convert light energy into electrical signals, enabling high-quality digital image acquisition. The core function of this imager is to provide reliable and precise image data for various laboratory applications.

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9 protocols using ccd camera based imager

1

Quantification of NET Signaling Molecules

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The cells with different treatments were lysed in RIPA buffer (25 mM Tris–HCl pH 7.6, 150 mM NaCl, 1% NP-40, 1% sodium deoxycholate, and 0.1% SDS) containing a protease inhibitor cocktail (Roche Applied Science). Twenty micrograms of total protein from sEVs lysate were loaded and separated on a standard sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE) gel and transferred to a polyvinylidene difluoride (PVDF) membrane (Millipore, Billerica, MA, USA). The membranes were incubated with primary antibodies to the NETs signaling molecules, followed by peroxidase-conjugated secondary antibodies. The results were detected using a charge-coupled device (CCD) camera-based imager (GE Healthcare Life Sciences, Chicago, IL, USA) after membrane incubation with enhanced chemiluminescence (ECL) substrates (Millipore). The protein levels of NETs signaling molecules were normalized to β-actin.
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2

Protein Expression Analysis of Engineered Breast Cancer Cells

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E0771, E0771-dCas9-VP64, and E0771-dCas9-VP64-MPH cells in 6-well plates were washed twice with ice cold PBS before lysis with 1× RIPA buffer which was kept on ice for 15 min. Cell lysates were centrifuged at 12,000 g for 15 min at 4°C and protein-containing supernatant was collected. Protein concentration was measured using a standard Bradford assay (BioRad) and 20 μg of protein in each sample were loaded into SDS-PAGE gel. After electrophoresis, proteins separated in gel were transferred into nitrocellulose membranes. Membranes were blocked at room temperature for 1 h using 5% skim-milk in TBST, followed by the incubation with primary antibody in 4°C overnight. After washing three times with TBST, horseradish peroxidase (HRP)-conjugated secondary antibody was added and incubated at room temperature for 30 – 60 min. The chemiluminescent substrate (Clarity Western ECL Substrate, Bio-Rad) was added on top of blot membrane according to the manufacturer’s instructions. The signals were captured using a CCD camera-based imager (GE Healthcare).
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3

Protein Expression Analysis of Exosomes

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The cells with different treatments were lysed in RIPA buffer (25 mM Tris–HCl pH 7.6, 150 mM NaCl, 1% NP-40, 1% sodium deoxycholate and 0.1% SDS) containing a protease inhibitor cocktail (Complete, Roche, Germany). Twenty micrograms of total protein from exosome lysate were loaded and separated on a standard sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE) gel and transferred to a polyvinylidene difluoride (PVDF) membrane (Millipore, USA). The membranes were incubated with primary antibodies, followed by peroxidase-conjugated secondary antibodies. The results were detected using a charge-coupled device (CCD) camera-based imager (GE Healthcare Life Sciences, USA) after membrane incubation with enhanced chemiluminescence (ECL) substrates (Millipore, USA). The levels of specific protein were normalized to β-actin. The ImageJ software was used for image acquisition and densitometric analysis of the immunoblots. All results were obtained in three independent experiments and the data are presented as the mean ± SD. An unpaired, two-tailed Student’s t-test was performed for between-group comparisons using GraphPad Prism software version 8. All results of densitometric analysis are presented in Additional file 2.
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4

Western Blot Protein Analysis Protocol

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The cells with different treatments were lysed in a radio-immunoprecipitation assay (RIPA) buffer (25 mM Tris-HCl [pH 7.6], 150 mM NaCl, 1% NP-40, 1% sodium deoxycholate, and 0.1% SDS) that contained a protease inhibitor cocktail (Complete, Roche, Germany). 20 μg of total protein from an exosome lysate were loaded and separated on a standard sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE) gel and were transferred to a polyvinylidene difluoride (PVDF) membrane (Millipore, USA). The membranes were incubated with primary antibodies, and this was followed by peroxidase-conjugated secondary antibodies. The results were detected using a charge-coupled device (CCD) camera-based imager (GE Healthcare Life Sciences) after membrane incubation with enhanced chemiluminescence (ECL) substrates (Millipore).
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5

Western Blot Protein Analysis

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Cells in 6-well plate or 10-cm dish were washed twice with ice cold PBS. The cells were then lysed with 1× RIPA buffer on ice for 15 min, or nuclear protein purification using nuclear extraction kit (Abcam). Cell lysates were centrifuged at 12,000 g for 15 min at 4°C and protein-containing supernatant was collected. Protein concentration was measured using a BCA assay (Abcam) and 20 μg of protein in each sample were loaded into SDS-PAGE gel. After electrophoresis, proteins separated in gel were transferred into nitrocellulose membranes. Membranes were blocked at room temperature for 1 h using 5% skim-milk in TBST, followed by the incubation with primary antibody in 4°C overnight. After washing three times with TBST, horseradish peroxidase (HRP)-conjugated secondary antibody was added and incubated at room temperature for 30 – 60 min. The chemiluminescent substrate (Clarity Western ECL Substrate, Bio-Rad) was added on top of blot membrane according to the manufacturer’s instructions. The signals were captured using a CCD camera-based imager (GE Healthcare).
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6

Protein Expression Analysis of Engineered Breast Cancer Cells

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E0771, E0771-dCas9-VP64, and E0771-dCas9-VP64-MPH cells in 6-well plates were washed twice with ice cold PBS before lysis with 1× RIPA buffer which was kept on ice for 15 min. Cell lysates were centrifuged at 12,000 g for 15 min at 4°C and protein-containing supernatant was collected. Protein concentration was measured using a standard Bradford assay (BioRad) and 20 μg of protein in each sample were loaded into SDS-PAGE gel. After electrophoresis, proteins separated in gel were transferred into nitrocellulose membranes. Membranes were blocked at room temperature for 1 h using 5% skim-milk in TBST, followed by the incubation with primary antibody in 4°C overnight. After washing three times with TBST, horseradish peroxidase (HRP)-conjugated secondary antibody was added and incubated at room temperature for 30 – 60 min. The chemiluminescent substrate (Clarity Western ECL Substrate, Bio-Rad) was added on top of blot membrane according to the manufacturer’s instructions. The signals were captured using a CCD camera-based imager (GE Healthcare).
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7

Western Blot Analysis of Exosome Lysates

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The cells with different treatments were lysed in RIPA buffer (25 mM Tris-HCl, pH 7.6, 150 mM NaCl, 1% NP-40, 1% sodium deoxycholate, and 0.1% sodium dodecyl sulfate [SDS]) containing a protease inhibitor cocktail (Complete, Roche, Germany). Twenty micrograms of total protein from exosome lysate was loaded and separated on a standard SDS-polyacrylamide gel electrophoresis (PAGE) gel and transferred to a polyvinylidene difluoride (PVDF) membrane (Millipore, USA). The membranes were incubated with primary antibodies, followed by peroxidase-conjugated secondary antibodies. The results were detected using a charge-coupled device (CCD) camera-based imager (GE Healthcare Life Sciences, USA) after membrane incubation with enhanced-chemiluminescence (ECL) substrates (Millipore, USA).
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8

Immunoblotting Analysis of Exosomal Proteins

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The cells with different treatments were lysed in RIPA buffer (25 mM Tris-HCl pH 7.6, 150 mM NaCl, 1% NP-40, 1% sodium deoxycholate, and 0.1% SDS) containing a protease inhibitor cocktail (Complete, Roche). Twenty micrograms of total protein from exosome lysate were loaded and separated on a standard sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE) gel and transferred to a polyvinylidene difluoride (PVDF) membrane (Millipore). The membranes were incubated with primary antibodies, followed by peroxidase-conjugated secondary antibodies. The results were detected using a charge-coupled device (CCD) camera-based imager (GE Healthcare Life Sciences) after membrane incubation with enhanced chemiluminescence (ECL) substrates (Millipore). The levels of specific protein were normalized to β-actin. ImageJ software was used for image acquisition and densitometric analysis of the immunoblots. All results were obtained in 3 independent experiments, and the data is presented as the mean ± SD. An unpaired, two-tailed Student's t test was performed for between-group comparisons using GraphPad Prism software version 8. All results of densitometric analysis were presented in in Supplementary File 1.
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9

Extracellular Vesicle Protein Analysis

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EVs, MSCs and MSC-CM were lysed by RIPA buffer containing Halt Protease
Inhibitor Cocktail (ThermoFisher Scientific). The protein extracts were
electrophoresed and the gel was transferred to a nitrocellulose membrane. The
primary antibodies against CD63 (System Biosciences), Flotillin1 and GM130 (Cell
Signaling Technology), and goat anti-rabbit IgG secondary antibody were
employed. The images were detected by GE CCD camera-based imager.
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