Structured Review

GE Healthcare imagequant las4000
Functional activity of miR-124a and miR-181a on target genes. (A) Schematic image of miR-124a putative target site in mouse Dlx5 3′-UTR and alignment of mir-124a with wild-type (WT) and mutant (MUT) 3′-UTR regions of Dlx5 showing complementary pairing. Mutated nucleotides are underlined. These seed regions are evolutionally well-conserved among higher vertebrates. (B) Schematic image of miR-181 putative target site in the mouse Msx2 mRNA 3′-UTR and alignment of miR-181a with wild-type (WT) and mutant (MUT) 3′-UTR regions of Msx2. Complementary pairing between miR-181a and Msx2 is shown. Mutated 3′-UTR nucleotides are underlined. These seed regions are evolutionally well-conserved among higher vertebrates. (C) MC3T3 cells were cotransfected with luciferase reporters carrying wild-type Dlx5 3′-UTR or mutated Dlx5 3′-UTR, phRL-null (Renilla plasmid), and 50 nM RNA oligonucleotides of mir-Control (mir-C) or miR-124a. The effects of miR-124a and control miRNAs on reporter constructs after 36 h are shown. The ratio of reporter (Firefly) to control phRL-null plasmid (Renilla) in relative luminescence units is plotted. Values represent means ± SD of 3 separate wells. (D) Functional activity of the luciferase reporter plasmid carrying wild-type or mutated Msx2 3′-UTR was assessed as described above ( Fig. 4B ). Values represent means ± SD of 3 separate wells. (E) miR-124a directly targets and regulates Dlx5 and inhibits osteoblastogenesis. MC3T3 osteoblast cells were transfected with miR-124a, miRNA-Control, or transfection reagent only (Mock) at the indicated concentrations. Western blots for Dlx5 and actin (as control) were performed on total cell lysates collected at 48 h. The bands of Dlx5, Msx2 and actin were detected by <t>ImageQuant</t> <t>LAS4000</t> (GE Healthcare UK Ltd, Little Chalfont, UK), and the intensity of bands was measured by ImageJ( http://rsb.info.nih.gov/ij/ ). Values represent means ± SD of 3 separate wells. *P
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Images

1) Product Images from "Involvement of MicroRNAs in Regulation of Osteoblastic Differentiation in Mouse Induced Pluripotent Stem Cells"

Article Title: Involvement of MicroRNAs in Regulation of Osteoblastic Differentiation in Mouse Induced Pluripotent Stem Cells

Journal: PLoS ONE

doi: 10.1371/journal.pone.0043800

Functional activity of miR-124a and miR-181a on target genes. (A) Schematic image of miR-124a putative target site in mouse Dlx5 3′-UTR and alignment of mir-124a with wild-type (WT) and mutant (MUT) 3′-UTR regions of Dlx5 showing complementary pairing. Mutated nucleotides are underlined. These seed regions are evolutionally well-conserved among higher vertebrates. (B) Schematic image of miR-181 putative target site in the mouse Msx2 mRNA 3′-UTR and alignment of miR-181a with wild-type (WT) and mutant (MUT) 3′-UTR regions of Msx2. Complementary pairing between miR-181a and Msx2 is shown. Mutated 3′-UTR nucleotides are underlined. These seed regions are evolutionally well-conserved among higher vertebrates. (C) MC3T3 cells were cotransfected with luciferase reporters carrying wild-type Dlx5 3′-UTR or mutated Dlx5 3′-UTR, phRL-null (Renilla plasmid), and 50 nM RNA oligonucleotides of mir-Control (mir-C) or miR-124a. The effects of miR-124a and control miRNAs on reporter constructs after 36 h are shown. The ratio of reporter (Firefly) to control phRL-null plasmid (Renilla) in relative luminescence units is plotted. Values represent means ± SD of 3 separate wells. (D) Functional activity of the luciferase reporter plasmid carrying wild-type or mutated Msx2 3′-UTR was assessed as described above ( Fig. 4B ). Values represent means ± SD of 3 separate wells. (E) miR-124a directly targets and regulates Dlx5 and inhibits osteoblastogenesis. MC3T3 osteoblast cells were transfected with miR-124a, miRNA-Control, or transfection reagent only (Mock) at the indicated concentrations. Western blots for Dlx5 and actin (as control) were performed on total cell lysates collected at 48 h. The bands of Dlx5, Msx2 and actin were detected by ImageQuant LAS4000 (GE Healthcare UK Ltd, Little Chalfont, UK), and the intensity of bands was measured by ImageJ( http://rsb.info.nih.gov/ij/ ). Values represent means ± SD of 3 separate wells. *P
Figure Legend Snippet: Functional activity of miR-124a and miR-181a on target genes. (A) Schematic image of miR-124a putative target site in mouse Dlx5 3′-UTR and alignment of mir-124a with wild-type (WT) and mutant (MUT) 3′-UTR regions of Dlx5 showing complementary pairing. Mutated nucleotides are underlined. These seed regions are evolutionally well-conserved among higher vertebrates. (B) Schematic image of miR-181 putative target site in the mouse Msx2 mRNA 3′-UTR and alignment of miR-181a with wild-type (WT) and mutant (MUT) 3′-UTR regions of Msx2. Complementary pairing between miR-181a and Msx2 is shown. Mutated 3′-UTR nucleotides are underlined. These seed regions are evolutionally well-conserved among higher vertebrates. (C) MC3T3 cells were cotransfected with luciferase reporters carrying wild-type Dlx5 3′-UTR or mutated Dlx5 3′-UTR, phRL-null (Renilla plasmid), and 50 nM RNA oligonucleotides of mir-Control (mir-C) or miR-124a. The effects of miR-124a and control miRNAs on reporter constructs after 36 h are shown. The ratio of reporter (Firefly) to control phRL-null plasmid (Renilla) in relative luminescence units is plotted. Values represent means ± SD of 3 separate wells. (D) Functional activity of the luciferase reporter plasmid carrying wild-type or mutated Msx2 3′-UTR was assessed as described above ( Fig. 4B ). Values represent means ± SD of 3 separate wells. (E) miR-124a directly targets and regulates Dlx5 and inhibits osteoblastogenesis. MC3T3 osteoblast cells were transfected with miR-124a, miRNA-Control, or transfection reagent only (Mock) at the indicated concentrations. Western blots for Dlx5 and actin (as control) were performed on total cell lysates collected at 48 h. The bands of Dlx5, Msx2 and actin were detected by ImageQuant LAS4000 (GE Healthcare UK Ltd, Little Chalfont, UK), and the intensity of bands was measured by ImageJ( http://rsb.info.nih.gov/ij/ ). Values represent means ± SD of 3 separate wells. *P

Techniques Used: Functional Assay, Activity Assay, Mutagenesis, Luciferase, Plasmid Preparation, Construct, Transfection, Western Blot

2) Product Images from "Mesoporous silica nanoparticles trigger mitophagy in endothelial cells and perturb neuronal network activity in a size- and time-dependent manner"

Article Title: Mesoporous silica nanoparticles trigger mitophagy in endothelial cells and perturb neuronal network activity in a size- and time-dependent manner

Journal: International Journal of Nanomedicine

doi: 10.2147/IJN.S127663

Effect of MSNPs on cell autophagy assessed by measuring the production of LC3 mediator. Notes: HUVECs were incubated with different concentrations (0.05–1 mg/mL) of MSNPs of 250 nm ( A , C ) or of 30 ( B , D ) for 2 or 24 hours. LC3 production was analyzed by immunoblotting followed by enhanced chemiluminescence detection, and representative blots are shown ( A , B ). The intensity of chemiluminescent spots was semiquantitatively estimated by ImageQuant LAS4000, and expressed as the ratio between the intensity of the spot of interest and the intensity of β-actin bands for cell lysates ( C , D ). Data are mean ± standard error of three separate experiments performed in triplicate. Results were compared by Student’s t -test. ** P
Figure Legend Snippet: Effect of MSNPs on cell autophagy assessed by measuring the production of LC3 mediator. Notes: HUVECs were incubated with different concentrations (0.05–1 mg/mL) of MSNPs of 250 nm ( A , C ) or of 30 ( B , D ) for 2 or 24 hours. LC3 production was analyzed by immunoblotting followed by enhanced chemiluminescence detection, and representative blots are shown ( A , B ). The intensity of chemiluminescent spots was semiquantitatively estimated by ImageQuant LAS4000, and expressed as the ratio between the intensity of the spot of interest and the intensity of β-actin bands for cell lysates ( C , D ). Data are mean ± standard error of three separate experiments performed in triplicate. Results were compared by Student’s t -test. ** P

Techniques Used: Incubation

3) Product Images from "Re-endothelialization of rat lung scaffolds through passive, gravity-driven seeding of segment-specific pulmonary endothelial cells"

Article Title: Re-endothelialization of rat lung scaffolds through passive, gravity-driven seeding of segment-specific pulmonary endothelial cells

Journal: Journal of tissue engineering and regenerative medicine

doi: 10.1002/term.2382

Culture time canbe extendedwithout detrimentto recellularizationbutretrograde seedingwas not as effective. (A) H E-stained tissue from seeded lungs cultured for 7 days showed similar histology to 3-day culture. (B) Seeding through the PVs delivered cells to the venous system, but cell aggregation and fragmentation were apparent. (C) Dual seeding of HUVECs through both the pulmonary arteries and veins led to similar histology as seeding through the PA alone. In dual-seeded lungs, HUVECs labelled with DiI or DiO dyes were imaged with an ImageQuant LAS4000 gel imager (D, E) immediately after seeding and (F, G) after 72 h of culture. Arterial (PA) seeding led to deposition of cells throughout the lung including (E) the distal ends. Cells seeded via the veins deposited cells in the proximal area of the lung. (F) After 3 days of bioreactor culture at 1 ml/ min, the lungs retained their initial seeding pattern. (G) Separation of the lung lobes confirmed that each lobe received cells. (H) Microscopy of fluorescently labelled cells seeded via the arteries (red) and veins (green) showed them in separate vascular compartments. In some portions, cells appeared to reside near alveoli, which may suggest a microvasculature/capillary location. Representative photos from one experiment.
Figure Legend Snippet: Culture time canbe extendedwithout detrimentto recellularizationbutretrograde seedingwas not as effective. (A) H E-stained tissue from seeded lungs cultured for 7 days showed similar histology to 3-day culture. (B) Seeding through the PVs delivered cells to the venous system, but cell aggregation and fragmentation were apparent. (C) Dual seeding of HUVECs through both the pulmonary arteries and veins led to similar histology as seeding through the PA alone. In dual-seeded lungs, HUVECs labelled with DiI or DiO dyes were imaged with an ImageQuant LAS4000 gel imager (D, E) immediately after seeding and (F, G) after 72 h of culture. Arterial (PA) seeding led to deposition of cells throughout the lung including (E) the distal ends. Cells seeded via the veins deposited cells in the proximal area of the lung. (F) After 3 days of bioreactor culture at 1 ml/ min, the lungs retained their initial seeding pattern. (G) Separation of the lung lobes confirmed that each lobe received cells. (H) Microscopy of fluorescently labelled cells seeded via the arteries (red) and veins (green) showed them in separate vascular compartments. In some portions, cells appeared to reside near alveoli, which may suggest a microvasculature/capillary location. Representative photos from one experiment.

Techniques Used: Staining, Cell Culture, Microscopy

Related Articles

Nucleic Acid Electrophoresis:

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Article Snippet: The protein (15 mg) was loaded into a 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred to nitrocellulose membranes (Amersham Pharmacia Biotech, Buckinghamshire, UK). .. The signals were detected by enhanced chemiluminescence reagents, and images were detected on ImageQuant LAS4000 (GE Healthcare, Piscataway, NJ, USA).

Transfection:

Article Title: Directed evolution of CRISPR-Cas9 to increase its specificity
Article Snippet: The WT-Cas9, Sniper-Cas9, eSpCas9(1.1), Cas9-HF1, evoCas9, and HypaCas9 proteins expressed in HEK293T cells after transfection were detected using western blotting. .. ImageQuant LAS4000 (GE healthcare) was used for digital imaging.

Article Title: Involvement of MicroRNAs in Regulation of Osteoblastic Differentiation in Mouse Induced Pluripotent Stem Cells
Article Snippet: Western Blot Analysis MC3T3-E1 osteo-progenitor cells were collected at 48 h after transfection of anti-miRNAs. .. The bands of Dlx5, Msx2 and actin were detected by ImageQuant LAS4000 (GE Healthcare UK Ltd, Little Chalfont, UK), and the intensity of bands was measured by ImageJ( http://rsb.info.nih.gov/ij/ ).

In Vitro:

Article Title: VPS13A and VPS13C are lipid transport proteins differentially localized at ER contact sites
Article Snippet: Paragraph title: In vitro lipid-binding assay ... NBD fluorescence was visualized using an ImageQuant LAS4000 (GE Healthcare), and total protein was visualized with Coomassie staining.

Fluorescence:

Article Title: VPS13A and VPS13C are lipid transport proteins differentially localized at ER contact sites
Article Snippet: .. NBD fluorescence was visualized using an ImageQuant LAS4000 (GE Healthcare), and total protein was visualized with Coomassie staining. .. In vitro lipid-transfer FRET assays Lipid-transfer reactions were performed in 50-µl volumes in 96-well plates (Nunc) containing a protein:lipid ratio of 1:400, with 0.125 µM protein, 25 µM donor liposomes (61% DOPC, 30% liver PE, 2% NBD-PS, 2% Rhodamine-PE, and 5% DGS-NTA [Ni]) and 25 µM acceptor liposomes (65% DOPC, 30% liver PE, and 5% PI(4,5P)2 ).

Protease Inhibitor:

Article Title: CXCR4- and BCR-triggered integrin activation in B-cell chronic lymphocytic leukemia cells depends on JAK2-activated Bruton’s tyrosine kinase
Article Snippet: Immunoblotting Cells were lysed in ice-cold 1% NP-40 lysis buffer containing complete protease inhibitor cocktail (Roche). .. After incubation with JAK2 primary antibody and HRP-coupled secondary antibody (GE Healthcare Life Science), immunoreactive bands were visualized by ECL detection (EMD Millipore), acquired by using ImageQuant Las4000 (GE Healthcare Life Science) and quantified by densitometric analysis (Quantity One, Bio-Rad).

BIA-KA:

Article Title: Use of Yeast Spores for Microencapsulation of Enzymes
Article Snippet: Protein concentration was determined by a bicinchoninic acid (BCA) protein assay kit (Beyotime, Jiangsu, China). .. Signals were visualized by Clarity Western ECL Substrate (Bio-Rad, Shanghai, China), and images were obtained by using ImageQuant LAS4000 (GE Healthcare Bio-Science, Uppsala, Sweden).

Western Blot:

Article Title: Mesoporous silica nanoparticles trigger mitophagy in endothelial cells and perturb neuronal network activity in a size- and time-dependent manner
Article Snippet: Paragraph title: Evaluation of cellular autophagy and apoptosis by sodium dodecyl sulfate polyacrylamide-gel electrophoresis (SDS-PAGE)/Western blotting (WB) ... The intensity of chemiluminescent spots was semiquantitatively estimated with an ImageQuant LAS4000 (GE Healthcare, Little Chalfont, UK) and expressed as the ratio between the intensity of the spot of interest and the intensity of β-actin bands for cell lysates.

Article Title: Directed evolution of CRISPR-Cas9 to increase its specificity
Article Snippet: Paragraph title: Western blotting ... ImageQuant LAS4000 (GE healthcare) was used for digital imaging.

Article Title: Involvement of MicroRNAs in Regulation of Osteoblastic Differentiation in Mouse Induced Pluripotent Stem Cells
Article Snippet: Paragraph title: Western Blot Analysis ... The bands of Dlx5, Msx2 and actin were detected by ImageQuant LAS4000 (GE Healthcare UK Ltd, Little Chalfont, UK), and the intensity of bands was measured by ImageJ( http://rsb.info.nih.gov/ij/ ).

Article Title: 6-sialyllactose ameliorates dihydrotestosterone-induced benign prostatic hyperplasia through suppressing VEGF-mediated angiogenesis
Article Snippet: Paragraph title: Western blot analysis ... To detect target proteins, the membrane was examined with a Pierce ECL plus (ThermoFischer Scientific) using ImageQuant LAS4000 (GE Healthcare, Pittsburgh, PA).

Article Title: Antidiabetic Drug Metformin Protects Neuronal Cells against Quinolinic Acid-Induced Excitotoxicity by Decreasing Intracellular Calcium
Article Snippet: Paragraph title: 5. Western blot assay ... The signals were detected by enhanced chemiluminescence reagents, and images were detected on ImageQuant LAS4000 (GE Healthcare, Piscataway, NJ, USA).

Article Title: Use of Yeast Spores for Microencapsulation of Enzymes
Article Snippet: .. Signals were visualized by Clarity Western ECL Substrate (Bio-Rad, Shanghai, China), and images were obtained by using ImageQuant LAS4000 (GE Healthcare Bio-Science, Uppsala, Sweden). ..

Article Title: Novel 1, 3-N, O-Spiroheterocyclic compounds inhibit heparanase activity and enhance nedaplatin-induced cytotoxicity in cervical cancer cells
Article Snippet: Paragraph title: Western blotting ... After consecutive washes, the membranes were incubated with secondary antibody (diluted 1:5000) at room temperature for 1 h. Then, the membranes were developed with enhanced chemiluminescence and imaged with ImageQuant LAS4000 (GE Healthcare Life Sciences, Pittsburgh, PA).

Article Title: Human mesenchymal stem cell-engineered hepatic cell sheets accelerate liver regeneration in mice
Article Snippet: Paragraph title: Western blot analysis of cell lysates ... The bands were detected by ImageQuant LAS4000 (GE Healthcare UK Ltd, Little Chalfont, UK).

Article Title: Involvement of CTCF in transcription regulation of EGR1 at early G1 phase as an architecture factor
Article Snippet: Paragraph title: Cellular extraction and western blot analysis ... The signal was detected by Chemi-Lumi One L (Nacalai) and ImageQuant LAS4000 (GE Healthcare Life Sciences).

Immunolabeling:

Article Title: Resveratrol reverses the Warburg effect by targeting the pyruvate dehydrogenase complex in colon cancer cells
Article Snippet: Immunoreactive bands were quantified using an ImageQuant LAS4000 (GE Healthcare) and the results analyzed by ImageJ (National Institutes of Health). .. The immunolabeled proteins were detected using an Odyssey Infrared Imaging System (LI-COR) and the results were analyzed with the Odyssey Application Software to obtain the integrated intensities.

Blocking Assay:

Article Title: Novel 1, 3-N, O-Spiroheterocyclic compounds inhibit heparanase activity and enhance nedaplatin-induced cytotoxicity in cervical cancer cells
Article Snippet: After blocking in non-fat milk solution at room temperature, the membranes were incubated with diluted primary antibodies against either proteins of interest or β-actin overnight at 4°C. .. After consecutive washes, the membranes were incubated with secondary antibody (diluted 1:5000) at room temperature for 1 h. Then, the membranes were developed with enhanced chemiluminescence and imaged with ImageQuant LAS4000 (GE Healthcare Life Sciences, Pittsburgh, PA).

Purification:

Article Title: VPS13A and VPS13C are lipid transport proteins differentially localized at ER contact sites
Article Snippet: In vitro lipid-binding assay 1 µl of either NBD-labeled PE, PS, PA, cholesterol, or ceramide (1 mg/ml in methanol) was incubated with 19 µl purified Vps13α or Vps13crystal (2 mg/ml) for either 30 min at 37°C or 2 h at 4°C. .. NBD fluorescence was visualized using an ImageQuant LAS4000 (GE Healthcare), and total protein was visualized with Coomassie staining.

Enzyme-linked Immunosorbent Assay:

Article Title: Painful, degenerating intervertebral discs up-regulate neurite sprouting and CGRP through nociceptive factors
Article Snippet: Duplicate 100 μl samples of each conditioned medium were incubated in ELISA plates overnight at 4°C. .. Arrays were imaged with the provided enhanced chemiluminescence kit using an ImageQuant LAS4000 (GE Healthcare, Baie d'Urfe, QC, Canada).

Concentration Assay:

Article Title: Painful, degenerating intervertebral discs up-regulate neurite sprouting and CGRP through nociceptive factors
Article Snippet: Conditioned media analysis The concentration of TNF-α (ELH-TNFALPHA-001; RayBiotech, Norcross, GA, USA), NGF (ELH-BNGF-001; RayBiotech), BDNF (ELH-BDNF-001; RayBiotech) and IL-1β (ELH-IL1BETA-001; RayBiotech) in the culture medium was quantified using ELISA according to manufacturers' instructions. .. Arrays were imaged with the provided enhanced chemiluminescence kit using an ImageQuant LAS4000 (GE Healthcare, Baie d'Urfe, QC, Canada).

Article Title: Novel 1, 3-N, O-Spiroheterocyclic compounds inhibit heparanase activity and enhance nedaplatin-induced cytotoxicity in cervical cancer cells
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Incubation:

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Article Title: Painful, degenerating intervertebral discs up-regulate neurite sprouting and CGRP through nociceptive factors
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Article Title: Neuronal Release of Cytokine IL-3 Triggered by Mechanosensitive Autostimulation of the P2X7 Receptor Is Neuroprotective
Article Snippet: Briefly, the experimental samples were first incubated with biotinylated detection antibodies and then incubated on nitrocellulose membranes spotted with 29 different anti-cytokine antibodies in duplicate. .. The production of light corresponding to levels of bound cytokine was determined with ImageQuant LAS4000 and the intensity of each spot was measured using ImageQuant TL analysis software (all GE Healthcare).

Article Title: 6-sialyllactose ameliorates dihydrotestosterone-induced benign prostatic hyperplasia through suppressing VEGF-mediated angiogenesis
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Article Title: Antidiabetic Drug Metformin Protects Neuronal Cells against Quinolinic Acid-Induced Excitotoxicity by Decreasing Intracellular Calcium
Article Snippet: The membranes were incubated with specific antibodies for ERK1/2, p-ERK1/2, p38, p-p38, JNK, p-JNK, or GAPDH. .. The signals were detected by enhanced chemiluminescence reagents, and images were detected on ImageQuant LAS4000 (GE Healthcare, Piscataway, NJ, USA).

Article Title: VPS13A and VPS13C are lipid transport proteins differentially localized at ER contact sites
Article Snippet: In vitro lipid-binding assay 1 µl of either NBD-labeled PE, PS, PA, cholesterol, or ceramide (1 mg/ml in methanol) was incubated with 19 µl purified Vps13α or Vps13crystal (2 mg/ml) for either 30 min at 37°C or 2 h at 4°C. .. NBD fluorescence was visualized using an ImageQuant LAS4000 (GE Healthcare), and total protein was visualized with Coomassie staining.

Article Title: Novel 1, 3-N, O-Spiroheterocyclic compounds inhibit heparanase activity and enhance nedaplatin-induced cytotoxicity in cervical cancer cells
Article Snippet: .. After consecutive washes, the membranes were incubated with secondary antibody (diluted 1:5000) at room temperature for 1 h. Then, the membranes were developed with enhanced chemiluminescence and imaged with ImageQuant LAS4000 (GE Healthcare Life Sciences, Pittsburgh, PA). ..

Article Title: CXCR4- and BCR-triggered integrin activation in B-cell chronic lymphocytic leukemia cells depends on JAK2-activated Bruton’s tyrosine kinase
Article Snippet: .. After incubation with JAK2 primary antibody and HRP-coupled secondary antibody (GE Healthcare Life Science), immunoreactive bands were visualized by ECL detection (EMD Millipore), acquired by using ImageQuant Las4000 (GE Healthcare Life Science) and quantified by densitometric analysis (Quantity One, Bio-Rad). ..

Article Title: Re-endothelialization of rat lung scaffolds through passive, gravity-driven seeding of segment-specific pulmonary endothelial cells
Article Snippet: Protected from direct light, 5 μL of Vybrant Dil or DiO cell-labelling solution was added per ml of cell suspension and incubated for 20 min at 37 °C in a tissue culture incubator. .. Cells were visualized immediately after seeding, as well as 72 h after seeding, using an ImageQuant LAS4000 (GE Healthcare Life Sciences, Pittsburgh, PA, USA).

Imaging:

Article Title: Resveratrol reverses the Warburg effect by targeting the pyruvate dehydrogenase complex in colon cancer cells
Article Snippet: Immunoreactive bands were quantified using an ImageQuant LAS4000 (GE Healthcare) and the results analyzed by ImageJ (National Institutes of Health). .. The immunolabeled proteins were detected using an Odyssey Infrared Imaging System (LI-COR) and the results were analyzed with the Odyssey Application Software to obtain the integrated intensities.

Article Title: Directed evolution of CRISPR-Cas9 to increase its specificity
Article Snippet: .. ImageQuant LAS4000 (GE healthcare) was used for digital imaging. .. Genomic DNA was purified from HEK293T cells with a DNeasy Blood & Tissue Kit (Qiagen).

Polyacrylamide Gel Electrophoresis:

Article Title: Mesoporous silica nanoparticles trigger mitophagy in endothelial cells and perturb neuronal network activity in a size- and time-dependent manner
Article Snippet: Paragraph title: Evaluation of cellular autophagy and apoptosis by sodium dodecyl sulfate polyacrylamide-gel electrophoresis (SDS-PAGE)/Western blotting (WB) ... The intensity of chemiluminescent spots was semiquantitatively estimated with an ImageQuant LAS4000 (GE Healthcare, Little Chalfont, UK) and expressed as the ratio between the intensity of the spot of interest and the intensity of β-actin bands for cell lysates.

Article Title: Novel 1, 3-N, O-Spiroheterocyclic compounds inhibit heparanase activity and enhance nedaplatin-induced cytotoxicity in cervical cancer cells
Article Snippet: 30 μg of protein was separated by 10% polyacrylamide gel electrophoresis (PAGE) and transferred to polyvinylidene difluoride membranes. .. After consecutive washes, the membranes were incubated with secondary antibody (diluted 1:5000) at room temperature for 1 h. Then, the membranes were developed with enhanced chemiluminescence and imaged with ImageQuant LAS4000 (GE Healthcare Life Sciences, Pittsburgh, PA).

Protein Concentration:

Article Title: Antidiabetic Drug Metformin Protects Neuronal Cells against Quinolinic Acid-Induced Excitotoxicity by Decreasing Intracellular Calcium
Article Snippet: Briefly, the cells were lysed in a lysis buffer and the protein concentration was determined with the Quant-iT™ assay kit (Molecular Probes, Eugene, OR, USA). .. The signals were detected by enhanced chemiluminescence reagents, and images were detected on ImageQuant LAS4000 (GE Healthcare, Piscataway, NJ, USA).

Article Title: Use of Yeast Spores for Microencapsulation of Enzymes
Article Snippet: Protein concentration was determined by a bicinchoninic acid (BCA) protein assay kit (Beyotime, Jiangsu, China). .. Signals were visualized by Clarity Western ECL Substrate (Bio-Rad, Shanghai, China), and images were obtained by using ImageQuant LAS4000 (GE Healthcare Bio-Science, Uppsala, Sweden).

Staining:

Article Title: VPS13A and VPS13C are lipid transport proteins differentially localized at ER contact sites
Article Snippet: .. NBD fluorescence was visualized using an ImageQuant LAS4000 (GE Healthcare), and total protein was visualized with Coomassie staining. .. In vitro lipid-transfer FRET assays Lipid-transfer reactions were performed in 50-µl volumes in 96-well plates (Nunc) containing a protein:lipid ratio of 1:400, with 0.125 µM protein, 25 µM donor liposomes (61% DOPC, 30% liver PE, 2% NBD-PS, 2% Rhodamine-PE, and 5% DGS-NTA [Ni]) and 25 µM acceptor liposomes (65% DOPC, 30% liver PE, and 5% PI(4,5P)2 ).

Lysis:

Article Title: Involvement of MicroRNAs in Regulation of Osteoblastic Differentiation in Mouse Induced Pluripotent Stem Cells
Article Snippet: Cells were lysed in lysis buffer containing protease inhibitors and 30 µg aliquots were subjected to western blot analysis using specific antibodies against Msx2 (Santa Cruz Biotechnology, California, USA), Dlx5 (Proteintech Group, Inc. Illinois, USA), and actin (Santa Cruz). .. The bands of Dlx5, Msx2 and actin were detected by ImageQuant LAS4000 (GE Healthcare UK Ltd, Little Chalfont, UK), and the intensity of bands was measured by ImageJ( http://rsb.info.nih.gov/ij/ ).

Article Title: 6-sialyllactose ameliorates dihydrotestosterone-induced benign prostatic hyperplasia through suppressing VEGF-mediated angiogenesis
Article Snippet: Western blot analysis Total proteins from HUVECs using 1% NP-40 lysis buffer. .. To detect target proteins, the membrane was examined with a Pierce ECL plus (ThermoFischer Scientific) using ImageQuant LAS4000 (GE Healthcare, Pittsburgh, PA).

Article Title: Antidiabetic Drug Metformin Protects Neuronal Cells against Quinolinic Acid-Induced Excitotoxicity by Decreasing Intracellular Calcium
Article Snippet: Briefly, the cells were lysed in a lysis buffer and the protein concentration was determined with the Quant-iT™ assay kit (Molecular Probes, Eugene, OR, USA). .. The signals were detected by enhanced chemiluminescence reagents, and images were detected on ImageQuant LAS4000 (GE Healthcare, Piscataway, NJ, USA).

Article Title: Novel 1, 3-N, O-Spiroheterocyclic compounds inhibit heparanase activity and enhance nedaplatin-induced cytotoxicity in cervical cancer cells
Article Snippet: Western blotting Cells were washed with precooled PBS after being treated with inhibitor for 48 hours, and cell lysis solution was added to extract protein. .. After consecutive washes, the membranes were incubated with secondary antibody (diluted 1:5000) at room temperature for 1 h. Then, the membranes were developed with enhanced chemiluminescence and imaged with ImageQuant LAS4000 (GE Healthcare Life Sciences, Pittsburgh, PA).

Article Title: CXCR4- and BCR-triggered integrin activation in B-cell chronic lymphocytic leukemia cells depends on JAK2-activated Bruton’s tyrosine kinase
Article Snippet: Immunoblotting Cells were lysed in ice-cold 1% NP-40 lysis buffer containing complete protease inhibitor cocktail (Roche). .. After incubation with JAK2 primary antibody and HRP-coupled secondary antibody (GE Healthcare Life Science), immunoreactive bands were visualized by ECL detection (EMD Millipore), acquired by using ImageQuant Las4000 (GE Healthcare Life Science) and quantified by densitometric analysis (Quantity One, Bio-Rad).

Article Title: Human mesenchymal stem cell-engineered hepatic cell sheets accelerate liver regeneration in mice
Article Snippet: Western blot analysis of cell lysates Ten micrograms of the cell lysis proteins were subjected to western blot analysis. .. The bands were detected by ImageQuant LAS4000 (GE Healthcare UK Ltd, Little Chalfont, UK).

Clear Native PAGE:

Article Title: VPS13A and VPS13C are lipid transport proteins differentially localized at ER contact sites
Article Snippet: Samples were visualized on 10% native PAGE gels. .. NBD fluorescence was visualized using an ImageQuant LAS4000 (GE Healthcare), and total protein was visualized with Coomassie staining.

SDS Page:

Article Title: Mesoporous silica nanoparticles trigger mitophagy in endothelial cells and perturb neuronal network activity in a size- and time-dependent manner
Article Snippet: After being boiled for 5 minutes at 100°C, an aliquot of each sample containing 15 µg total proteins was subjected to SDS-PAGE using precast NuPAGE 4%–12% gel (Thermo Fisher Scientific). .. The intensity of chemiluminescent spots was semiquantitatively estimated with an ImageQuant LAS4000 (GE Healthcare, Little Chalfont, UK) and expressed as the ratio between the intensity of the spot of interest and the intensity of β-actin bands for cell lysates.

Article Title: Use of Yeast Spores for Microencapsulation of Enzymes
Article Snippet: The cell lysates were then centrifuged at 4,000 × g for 5 min, and 50 μg of the supernatants was subjected to SDS-PAGE (5% stacking gel and 10% separating gel). .. Signals were visualized by Clarity Western ECL Substrate (Bio-Rad, Shanghai, China), and images were obtained by using ImageQuant LAS4000 (GE Healthcare Bio-Science, Uppsala, Sweden).

Software:

Article Title: Painful, degenerating intervertebral discs up-regulate neurite sprouting and CGRP through nociceptive factors
Article Snippet: Colorimetric absorbance was measured with a Tecan Infinite M200 PRO (Tecan, Männedorf, Switzerland) and analysed with i-control 1.9 software (Tecan). .. Arrays were imaged with the provided enhanced chemiluminescence kit using an ImageQuant LAS4000 (GE Healthcare, Baie d'Urfe, QC, Canada).

Article Title: Resveratrol reverses the Warburg effect by targeting the pyruvate dehydrogenase complex in colon cancer cells
Article Snippet: Immunoreactive bands were quantified using an ImageQuant LAS4000 (GE Healthcare) and the results analyzed by ImageJ (National Institutes of Health). .. The immunolabeled proteins were detected using an Odyssey Infrared Imaging System (LI-COR) and the results were analyzed with the Odyssey Application Software to obtain the integrated intensities.

Article Title: Neuronal Release of Cytokine IL-3 Triggered by Mechanosensitive Autostimulation of the P2X7 Receptor Is Neuroprotective
Article Snippet: .. The production of light corresponding to levels of bound cytokine was determined with ImageQuant LAS4000 and the intensity of each spot was measured using ImageQuant TL analysis software (all GE Healthcare). .. Measurement of IL-3 IL-3 concentrations were determined using the Rat IL-3 ELISA kit (#CSB-E07436r, Cusabio Biotech Co., College Park, MD, USA) by following the manufacturer’s instructions.

Article Title: Re-endothelialization of rat lung scaffolds through passive, gravity-driven seeding of segment-specific pulmonary endothelial cells
Article Snippet: Cells were visualized immediately after seeding, as well as 72 h after seeding, using an ImageQuant LAS4000 (GE Healthcare Life Sciences, Pittsburgh, PA, USA). .. Tissue sections (5 μm) were cut on a cryotome, mounted and coverslipped with Prolong Gold Antifade Reagent containing DAPI (Invitrogen Thermo Fisher Scientific, Waltham, MA, USA) before photographing on a Nikon Eclipse E800 fitted with a Qlmaging Retiga 2000R camera and Slidebook software.

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    GE Healthcare imagequanttm las400
    Imagequanttm Las400, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 92/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    GE Healthcare imagequanttm las4000 digital imaging system
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