opti mem reduced serum medium  (Thermo Fisher)


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    Name:
    Opti MEM I Reduced Serum Medium
    Description:
    Opti MEM I Reduced Serum Medium is an improved Minimal Essential Medium MEM that allows for a reduction of Fetal Bovine Serum supplementation by at least 50 with no change in cell growth rate or morphology Opti MEM I medium is also recommended for use with cationic lipid transfection reagents such as Lipofectamine reagent Opti MEM I medium can be used with a variety of suspension and adherent mammalian cells including Sp2 AE 1 CHO BHK 21 HEK and primary fibroblasts We offer a variety of Opti MEM I Reduced Serum Medium modifications for a range of cell culture applications This Opti MEM is modified as follows WithWithout• L glutamine• Phenol RedThe complete formulation is confidential For more information please contact Technical Services Using Opti MEM Reduced Serum MediumOpti MEM I medium is a unique medium that contains insulin transferrin hypoxanthine thymidine and trace elements These additional components allow for a reduction in serum supplementation by at least 50 Opti MEM I medium uses a sodium bicarbonate buffer system 2 4 g L and therefore requires a 5 10 CO2 environment to maintain physiological pH cGMP manufacturing and quality systemFor supply chain continuity we manufacture Opti MEM Reduced Serum Medium at two separate facilities located in Grand Island NY and Scotland UK Both sites are compliant with cGMP manufacturing requirements are certified to ISO 13485 and are registered with the FDA as medical device manufacturers
    Catalog Number:
    11058021
    Price:
    None
    Applications:
    Cell Culture|Mammalian Cell Culture|Plasmid Transfection|RNAi Transfection|Synthetic siRNA Transfection|shRNA & miR RNAi Plasmid Transfection|Transfection
    Category:
    Cell Culture Transfection Reagents
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    Structured Review

    Thermo Fisher opti mem reduced serum medium
    IGFBP7 inhibits the growth of AML cells. ( a ) Average expression of IGFBP7 measured using qRT-PCR, error bars show the S.D. of a duplicate. ( b ) Lentivirally transduced leukemic cell lines were analysed for the expression of IGFBP7 by immunoblotting of the lysates (upper panel) and the medium (middle panel). The expression of actin was used as a control for equal loading (lower panel). * indicates an aspecific background band. ( c ) Cell viability of Kasumi-1 cells transduced with the control (EF-1) or with the IGFBP7 overexpression vector were cultured under low-serum conditions (1%) for 72 h. Bars and error bars show the cell viability and S.D. of a triplicate. ( d ) AML cells were treated with various concentrations of rhIGFBP7 in <t>Opti-Mem</t> reduced serum medium with 1% <t>FCS</t> for 72 h. Bars represent the average and error bars represent the viability and S.D. of a triplicate. One-way ANOVA analysis was used with a post hoc Tukey test to calculate the P -value * P ≤0.05, ** P ≤0.01, *** P ≤0.001
    Opti MEM I Reduced Serum Medium is an improved Minimal Essential Medium MEM that allows for a reduction of Fetal Bovine Serum supplementation by at least 50 with no change in cell growth rate or morphology Opti MEM I medium is also recommended for use with cationic lipid transfection reagents such as Lipofectamine reagent Opti MEM I medium can be used with a variety of suspension and adherent mammalian cells including Sp2 AE 1 CHO BHK 21 HEK and primary fibroblasts We offer a variety of Opti MEM I Reduced Serum Medium modifications for a range of cell culture applications This Opti MEM is modified as follows WithWithout• L glutamine• Phenol RedThe complete formulation is confidential For more information please contact Technical Services Using Opti MEM Reduced Serum MediumOpti MEM I medium is a unique medium that contains insulin transferrin hypoxanthine thymidine and trace elements These additional components allow for a reduction in serum supplementation by at least 50 Opti MEM I medium uses a sodium bicarbonate buffer system 2 4 g L and therefore requires a 5 10 CO2 environment to maintain physiological pH cGMP manufacturing and quality systemFor supply chain continuity we manufacture Opti MEM Reduced Serum Medium at two separate facilities located in Grand Island NY and Scotland UK Both sites are compliant with cGMP manufacturing requirements are certified to ISO 13485 and are registered with the FDA as medical device manufacturers
    https://www.bioz.com/result/opti mem reduced serum medium/product/Thermo Fisher
    Average 99 stars, based on 715 article reviews
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    opti mem reduced serum medium - by Bioz Stars, 2020-07
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    Images

    1) Product Images from "IGFBP7 induces apoptosis of acute myeloid leukemia cells and synergizes with chemotherapy in suppression of leukemia cell survival"

    Article Title: IGFBP7 induces apoptosis of acute myeloid leukemia cells and synergizes with chemotherapy in suppression of leukemia cell survival

    Journal: Cell Death & Disease

    doi: 10.1038/cddis.2014.268

    IGFBP7 inhibits the growth of AML cells. ( a ) Average expression of IGFBP7 measured using qRT-PCR, error bars show the S.D. of a duplicate. ( b ) Lentivirally transduced leukemic cell lines were analysed for the expression of IGFBP7 by immunoblotting of the lysates (upper panel) and the medium (middle panel). The expression of actin was used as a control for equal loading (lower panel). * indicates an aspecific background band. ( c ) Cell viability of Kasumi-1 cells transduced with the control (EF-1) or with the IGFBP7 overexpression vector were cultured under low-serum conditions (1%) for 72 h. Bars and error bars show the cell viability and S.D. of a triplicate. ( d ) AML cells were treated with various concentrations of rhIGFBP7 in Opti-Mem reduced serum medium with 1% FCS for 72 h. Bars represent the average and error bars represent the viability and S.D. of a triplicate. One-way ANOVA analysis was used with a post hoc Tukey test to calculate the P -value * P ≤0.05, ** P ≤0.01, *** P ≤0.001
    Figure Legend Snippet: IGFBP7 inhibits the growth of AML cells. ( a ) Average expression of IGFBP7 measured using qRT-PCR, error bars show the S.D. of a duplicate. ( b ) Lentivirally transduced leukemic cell lines were analysed for the expression of IGFBP7 by immunoblotting of the lysates (upper panel) and the medium (middle panel). The expression of actin was used as a control for equal loading (lower panel). * indicates an aspecific background band. ( c ) Cell viability of Kasumi-1 cells transduced with the control (EF-1) or with the IGFBP7 overexpression vector were cultured under low-serum conditions (1%) for 72 h. Bars and error bars show the cell viability and S.D. of a triplicate. ( d ) AML cells were treated with various concentrations of rhIGFBP7 in Opti-Mem reduced serum medium with 1% FCS for 72 h. Bars represent the average and error bars represent the viability and S.D. of a triplicate. One-way ANOVA analysis was used with a post hoc Tukey test to calculate the P -value * P ≤0.05, ** P ≤0.01, *** P ≤0.001

    Techniques Used: Expressing, Quantitative RT-PCR, Transduction, Over Expression, Plasmid Preparation, Cell Culture

    2) Product Images from "Role of the ANKMY2-FKBP38 Axis in Regulation of the Sonic Hedgehog (Shh) Signaling Pathway *"

    Article Title: Role of the ANKMY2-FKBP38 Axis in Regulation of the Sonic Hedgehog (Shh) Signaling Pathway *

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M114.558635

    FKBP38 is a negative regulator of Shh signal transduction. A , confluent cultures of Fkbp38 +/+ or Fkbp38 −/− MEFs that had been deprived of serum for 24 h by incubation in Opti-MEM reduced serum medium were fixed and processed for immunofluorescence analysis with antibodies to γ-tubulin and to acetylated α-tubulin. Nuclei were also stained with Hoechst 33258, and the cells were then examined with a confocal microscope. Merged images are also shown; scale bars , 20 μm. Higher magnification views are boxed ; scale bars , 5 μm. B , Fkbp38 +/+ and Fkbp38 −/− MEFs were incubated in the absence or presence of Shh for 48 h, after which whole cell homogenates were prepared and subjected to immunoblot analysis ( IB ) with anti-Gli1, anti-FKBP38, anti-ANKMY2, and anti-HSP90. C , Fkbp38 +/+ and Fkbp38 −/− MEFs were incubated in the absence or presence of Shh for 24 h, after which total RNA was extracted from the cells and subjected to RT and real-time PCR analysis of Gli1 and Ptch1 mRNAs. Data are presented as relative -fold induction by Shh and are means ± S.D. from five independent experiments. **, p
    Figure Legend Snippet: FKBP38 is a negative regulator of Shh signal transduction. A , confluent cultures of Fkbp38 +/+ or Fkbp38 −/− MEFs that had been deprived of serum for 24 h by incubation in Opti-MEM reduced serum medium were fixed and processed for immunofluorescence analysis with antibodies to γ-tubulin and to acetylated α-tubulin. Nuclei were also stained with Hoechst 33258, and the cells were then examined with a confocal microscope. Merged images are also shown; scale bars , 20 μm. Higher magnification views are boxed ; scale bars , 5 μm. B , Fkbp38 +/+ and Fkbp38 −/− MEFs were incubated in the absence or presence of Shh for 48 h, after which whole cell homogenates were prepared and subjected to immunoblot analysis ( IB ) with anti-Gli1, anti-FKBP38, anti-ANKMY2, and anti-HSP90. C , Fkbp38 +/+ and Fkbp38 −/− MEFs were incubated in the absence or presence of Shh for 24 h, after which total RNA was extracted from the cells and subjected to RT and real-time PCR analysis of Gli1 and Ptch1 mRNAs. Data are presented as relative -fold induction by Shh and are means ± S.D. from five independent experiments. **, p

    Techniques Used: Transduction, Incubation, Immunofluorescence, Staining, Microscopy, Real-time Polymerase Chain Reaction

    FKBP38 regulates ANKMY2 localization. A , confluent cultures of Fkbp38 +/+ or Fkbp38 −/− MEFs that had been deprived of serum for 24 h by incubation in Opti-MEM reduced serum medium were fixed and processed for immunofluorescence analysis with antibodies to acetylated α-tubulin and to FKBP38. Nuclei were stained with Hoechst 33258, and the cells were then examined with a confocal microscope. Merged images are also shown; scale bars , 20 μm. Higher magnification views are boxed ; scale bar , 5 μm. B , confluent cultures of MEFs stably expressing EGFP-tagged human ANKMY2 were deprived of serum for 24 h, fixed, and processed for immunofluorescence analysis with antibodies to acetylated α-tubulin. Nuclei were stained with Hoechst 33258, and the cells were then examined with a confocal microscope. The fluorescence of EGFP-ANKMY2 was monitored directly. A merged image is also shown; scale bar , 20 μm. Higher magnification views are boxed ; scale bar , 5 μm. C , HeLa cells expressing 2×HA-tagged human FKBP38 and 3×FLAG-tagged mouse ANKMY2 or ANKMY2(ΔMYND) were fixed and processed for immunofluorescence staining with anti-FLAG and anti-HA. Nuclei were stained with Hoechst 33258, and the cells were then examined with a confocal microscope. Merged images are also shown; scale bars , 20 μm. Higher magnification views are boxed ; scale bars , 10 μm. An arrowhead indicates colocalization of ANKMY2 and FKBP38 signals; an arrow indicates that ANKMY2(ΔMYND) and FKBP38 signals are not colocalized. D , HeLa cells expressing 3×FLAG-tagged ANKMY2 and 2×HA-tagged FKBP38 or FKBP38(ΔN57) were fixed and processed for immunofluorescence staining with anti-HA and anti-FLAG. Nuclei were stained with Hoechst 33258, and the cells were then examined with a confocal microscope. Merged images are also shown; scale bars , 20 μm. Higher magnification views are boxed ; scale bars , 10 μm. An arrowhead indicates colocalization of ANKMY2 and FKBP38 signals; an arrow indicates that ANKMY2 and FKBP38(ΔN57) signals are not colocalized.
    Figure Legend Snippet: FKBP38 regulates ANKMY2 localization. A , confluent cultures of Fkbp38 +/+ or Fkbp38 −/− MEFs that had been deprived of serum for 24 h by incubation in Opti-MEM reduced serum medium were fixed and processed for immunofluorescence analysis with antibodies to acetylated α-tubulin and to FKBP38. Nuclei were stained with Hoechst 33258, and the cells were then examined with a confocal microscope. Merged images are also shown; scale bars , 20 μm. Higher magnification views are boxed ; scale bar , 5 μm. B , confluent cultures of MEFs stably expressing EGFP-tagged human ANKMY2 were deprived of serum for 24 h, fixed, and processed for immunofluorescence analysis with antibodies to acetylated α-tubulin. Nuclei were stained with Hoechst 33258, and the cells were then examined with a confocal microscope. The fluorescence of EGFP-ANKMY2 was monitored directly. A merged image is also shown; scale bar , 20 μm. Higher magnification views are boxed ; scale bar , 5 μm. C , HeLa cells expressing 2×HA-tagged human FKBP38 and 3×FLAG-tagged mouse ANKMY2 or ANKMY2(ΔMYND) were fixed and processed for immunofluorescence staining with anti-FLAG and anti-HA. Nuclei were stained with Hoechst 33258, and the cells were then examined with a confocal microscope. Merged images are also shown; scale bars , 20 μm. Higher magnification views are boxed ; scale bars , 10 μm. An arrowhead indicates colocalization of ANKMY2 and FKBP38 signals; an arrow indicates that ANKMY2(ΔMYND) and FKBP38 signals are not colocalized. D , HeLa cells expressing 3×FLAG-tagged ANKMY2 and 2×HA-tagged FKBP38 or FKBP38(ΔN57) were fixed and processed for immunofluorescence staining with anti-HA and anti-FLAG. Nuclei were stained with Hoechst 33258, and the cells were then examined with a confocal microscope. Merged images are also shown; scale bars , 20 μm. Higher magnification views are boxed ; scale bars , 10 μm. An arrowhead indicates colocalization of ANKMY2 and FKBP38 signals; an arrow indicates that ANKMY2 and FKBP38(ΔN57) signals are not colocalized.

    Techniques Used: Incubation, Immunofluorescence, Staining, Microscopy, Stable Transfection, Expressing, Fluorescence

    3) Product Images from "Acid ceramidase is upregulated in AML and represents a novel therapeutic target"

    Article Title: Acid ceramidase is upregulated in AML and represents a novel therapeutic target

    Journal: Oncotarget

    doi: 10.18632/oncotarget.13079

    S1P promotes survival and AC activity regulates sphingolipid profiles in AML cell lines A. Enhanced survival phenotype upon AC overexpression. Pro-apoptotic lipid levels decreased while pro-survival levels were significantly elevated in HL-60/AC cells relative to parental cells. B. Exogenous S1P increases viability in AML cell lines. Cell lines were incubated for 24 hours with 0.1 μM S1P or vehicle control in Opti-MEM reduced-serum (50%) medium. Viability assay results are normalized to cells incubated with medium containing 10% serum. C. Total ceramide levels and D. C 16 and C 24 ceramides in HL-60/VCR increased with LCL204 treatment (7.5 μM) for 12 hours. E. Exogenous ceramide decreased Mcl-1 expression. HL-60/VCR cells were treated with DMSO, C 16 ceramide (15 μM) or C 24 ceramide (2.5 μM) for 2 hours and 8 hours, then Mcl-1 and β-actin levels were determined by western blotting. F-G. LCL204 treatment altered sphingosine and S1P levels in human AML cell lines. Cells were incubated with either DMSO or LCL204 (7.5 μM) for 12h and (F) sphingosine and (G) S1P levels were quantified by mass spectrometry as described in the Materials and Method section. *, p
    Figure Legend Snippet: S1P promotes survival and AC activity regulates sphingolipid profiles in AML cell lines A. Enhanced survival phenotype upon AC overexpression. Pro-apoptotic lipid levels decreased while pro-survival levels were significantly elevated in HL-60/AC cells relative to parental cells. B. Exogenous S1P increases viability in AML cell lines. Cell lines were incubated for 24 hours with 0.1 μM S1P or vehicle control in Opti-MEM reduced-serum (50%) medium. Viability assay results are normalized to cells incubated with medium containing 10% serum. C. Total ceramide levels and D. C 16 and C 24 ceramides in HL-60/VCR increased with LCL204 treatment (7.5 μM) for 12 hours. E. Exogenous ceramide decreased Mcl-1 expression. HL-60/VCR cells were treated with DMSO, C 16 ceramide (15 μM) or C 24 ceramide (2.5 μM) for 2 hours and 8 hours, then Mcl-1 and β-actin levels were determined by western blotting. F-G. LCL204 treatment altered sphingosine and S1P levels in human AML cell lines. Cells were incubated with either DMSO or LCL204 (7.5 μM) for 12h and (F) sphingosine and (G) S1P levels were quantified by mass spectrometry as described in the Materials and Method section. *, p

    Techniques Used: Activity Assay, Over Expression, Incubation, Viability Assay, Expressing, Western Blot, Mass Spectrometry

    4) Product Images from "Ablation of Akt2 Induces Autophagy through Cell Cycle Arrest, the Downregulation of p70S6K, and the Deregulation of Mitochondria in MDA-MB231 Cells"

    Article Title: Ablation of Akt2 Induces Autophagy through Cell Cycle Arrest, the Downregulation of p70S6K, and the Deregulation of Mitochondria in MDA-MB231 Cells

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0014614

    Cdk2 and cyclin D were downregulated in cells transfected with Akt2-siRNA. ( A ) The relative level of Cdk2 was measured in a time course manner (0–120 h post-transfection) after cells were transfected with Akt2 siRNA (set #2). (Lower panel) The relative intensity of bands shown in the upper panel was normalized with the loading control and presented in a graph format. “No serum” denotes Opti-MEM transfection medium described in Materials and Methods . “Full” denotes culture medium containing 10% FBS (i.e., complete medium). ( B ) Analysis of Cdk2 by immunostaining in cells transfected with Akt2 siRNA (either oligo #1 or #2). The sampling times were 24 and 48 h post-transfection. Images shown are representative of three independent trials for each timepoint and each siRNA, respectively. ( C ) Akt2 ablation resulted in the downregulation of cyclin D, but not cyclin A and E. Western blot analysis of cyclin A, D and E of the cells transfected with Akt2 siRNA or scrambled siRNA was carried out at 0–120 h post-transfection. Note that the 0 h timepoint is incubated for 24 h in serum-free medium for both Akt2-siRNA and scrambled-siRNA treated cells. Data shown is representative of three independent experiments.
    Figure Legend Snippet: Cdk2 and cyclin D were downregulated in cells transfected with Akt2-siRNA. ( A ) The relative level of Cdk2 was measured in a time course manner (0–120 h post-transfection) after cells were transfected with Akt2 siRNA (set #2). (Lower panel) The relative intensity of bands shown in the upper panel was normalized with the loading control and presented in a graph format. “No serum” denotes Opti-MEM transfection medium described in Materials and Methods . “Full” denotes culture medium containing 10% FBS (i.e., complete medium). ( B ) Analysis of Cdk2 by immunostaining in cells transfected with Akt2 siRNA (either oligo #1 or #2). The sampling times were 24 and 48 h post-transfection. Images shown are representative of three independent trials for each timepoint and each siRNA, respectively. ( C ) Akt2 ablation resulted in the downregulation of cyclin D, but not cyclin A and E. Western blot analysis of cyclin A, D and E of the cells transfected with Akt2 siRNA or scrambled siRNA was carried out at 0–120 h post-transfection. Note that the 0 h timepoint is incubated for 24 h in serum-free medium for both Akt2-siRNA and scrambled-siRNA treated cells. Data shown is representative of three independent experiments.

    Techniques Used: Transfection, Immunostaining, Sampling, Western Blot, Incubation

    Akt2 ablation induced autophagy of mitochondria. ( A ) Akt2 ablation with siRNA resulted in an increase of LC3B-II. Western blotting was carried out with anti-LC3B antibody that recognizes both LC3B-I and LC3B-II. Cell extracts from Akt2-ablated or control cells (scrambled) were prepared at the timepoints indicated. “No” denotes no serum in the medium (i.e., Opti-MEM). Note the increased LC3B-II levels in the Akt2-ablated cells compared to the scrambled controls. ( B ) Immunofluorescent staining of LC3B was carried out at 96 h post-transfection. Red, green, and blue are MitoTracker (mitochondrial detection), LC3B (detection of LC3B, which is associated with autophagosomes), and DRAQ5 (nucleus detection), respectively. Akt2 siRNA and “Scrambled” denote MDA-MB231 cells transfected with Akt2 siRNA or scrambled siRNA, respectively. ( C ) Cells on a glass coverslip were transfected with Akt2 siRNA or scrambled siRNA, followed by triple-staining, first with MitoTracker Red, followed by acridine orange (AO) and monodansylcadaverine (MDC). Sampling timepoints after transfection are indicated left to panel.
    Figure Legend Snippet: Akt2 ablation induced autophagy of mitochondria. ( A ) Akt2 ablation with siRNA resulted in an increase of LC3B-II. Western blotting was carried out with anti-LC3B antibody that recognizes both LC3B-I and LC3B-II. Cell extracts from Akt2-ablated or control cells (scrambled) were prepared at the timepoints indicated. “No” denotes no serum in the medium (i.e., Opti-MEM). Note the increased LC3B-II levels in the Akt2-ablated cells compared to the scrambled controls. ( B ) Immunofluorescent staining of LC3B was carried out at 96 h post-transfection. Red, green, and blue are MitoTracker (mitochondrial detection), LC3B (detection of LC3B, which is associated with autophagosomes), and DRAQ5 (nucleus detection), respectively. Akt2 siRNA and “Scrambled” denote MDA-MB231 cells transfected with Akt2 siRNA or scrambled siRNA, respectively. ( C ) Cells on a glass coverslip were transfected with Akt2 siRNA or scrambled siRNA, followed by triple-staining, first with MitoTracker Red, followed by acridine orange (AO) and monodansylcadaverine (MDC). Sampling timepoints after transfection are indicated left to panel.

    Techniques Used: Western Blot, Staining, Transfection, Multiple Displacement Amplification, Sampling

    (A) Akt2 ablation caused an increase in the mitochondrial volume. Cells transfected with either Akt2 siRNA (i–vi) or scrambled siRNA (vii–xii) were immunostained with anti-Akt2 antibodies (Akt2) (green) or MitoTracker (red). Samples were taken at 0 or 48 h post-transfection as indicated. It should be noted that Akt2 is co-localized with mitochondria (ix, xii) as described previously [102] . Cells at 0 h were in the Opti-MEM (serum-free) for 24 h during transfection, and those at 48 h were maintained in the complete medium containing 10% FBS for 48 h after the transfection procedure was complete. Note that the 20 µm size bar of the scrambled samples is 1.3 fold bigger than that of Akt2-treated samples. ( B ) The relative surface area of the total mitochondria was compared to the total surface area of the cell (µm 2 ). The quantification of the cells surface area was carried out using software installed in the LSM 510 Meta laser scanning confocal microscope (Carl Zeiss). For the quantification, at least ten fields were analyzed for each sample. Error bars are standard errors. ( C ) The same as panel ( B ), except this data shows total cell area (µm 2 ).
    Figure Legend Snippet: (A) Akt2 ablation caused an increase in the mitochondrial volume. Cells transfected with either Akt2 siRNA (i–vi) or scrambled siRNA (vii–xii) were immunostained with anti-Akt2 antibodies (Akt2) (green) or MitoTracker (red). Samples were taken at 0 or 48 h post-transfection as indicated. It should be noted that Akt2 is co-localized with mitochondria (ix, xii) as described previously [102] . Cells at 0 h were in the Opti-MEM (serum-free) for 24 h during transfection, and those at 48 h were maintained in the complete medium containing 10% FBS for 48 h after the transfection procedure was complete. Note that the 20 µm size bar of the scrambled samples is 1.3 fold bigger than that of Akt2-treated samples. ( B ) The relative surface area of the total mitochondria was compared to the total surface area of the cell (µm 2 ). The quantification of the cells surface area was carried out using software installed in the LSM 510 Meta laser scanning confocal microscope (Carl Zeiss). For the quantification, at least ten fields were analyzed for each sample. Error bars are standard errors. ( C ) The same as panel ( B ), except this data shows total cell area (µm 2 ).

    Techniques Used: Transfection, Software, Microscopy

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    Article Snippet: .. Dulbecco's Phosphate-Buffered Saline without Ca2+ and Mg2+ (DPBS, Lonza, Catalog# 17-512F) DMEM, no glucose (Life Technologies, catalog# 11966-025) Sodium pyruvate (Life Technologies, catalog# 11360-070) D-(+)-Galactose (Sigma-Aldrich, catalog# G5388-100G) Paraformaldehyde (Electron Microscopy Science, catalog #15714) Sytox red (Life Technologies, catalog #34859) Goat serum (Rockland, catalog #D204-00-0050) 96-well Acoustic uClear Flat-Bottom Black-Wall Plates (Greiner, catalog #655090) 10X RIPA Lysis Buffer (0.5M Tris-HCl, pH 7.4, 1.5M NaCl, 2.5% deoxycholic acid, 10% NP-40, 10 mM EDTA) (Millipore, catalog #20-188) Sodium Dodecyl Sulfate (SDS, Sigma, catalog #L6026) Protease Inhibitor Cocktail (Sigma, catalog #P8340) Phosphatase Inhibitor Cocktail (Sigma, catalog #P0044) Halt Protease Inhibitor Cocktail, EDTA-free (Thermo Scientific, catalog #87785) M-PER® Mammalian Protein Extraction Reagent (Thermo Scientific, catalog #78501) NE-PER® Nuclear and Cytoplasmic Extraction Reagents (Thermo Scientific, catalog #28833) XDR Charge Separation Master Kit (ProteinSimple, catalog #CBS2001) pI Standard Ladder 3 (ProteinSimple, catalog #040-646) Premix G2, Pharmalyte pH 5-8 Separation Gradient (ProteinSimple, catalog #040-973) DMSO Inhibitor Mix (ProteinSimple, catalog #040-510) Wes™ 12-230 kDa Master Kit (ProteinSimple, catalog #PS-MK01 or PS-MK02) Bicine/CHAPS Lysis Buffer and Sample Diluent (ProteinSimple, catalog #040-764) Pierce BCA Protein Assay Kit (Thermo Scientific, catalog #23225) Lipofectamine RNAiMAX transfection reagent (Life Technologies, catalog #13778) Opti-MEM I Reduced Serum Medium (Life Technologies, catalog #31985-062) ON-TARGETplus SMARTpool Human ErbB2 siRNAs (GE Dharmacon, catalog #L-003126-00-0005) Non-targeting pool control siRNAs (GE Dharmacon, catalog #D-00180-10-05) Lapatinib (National Cancer Institute compound repository, NSC 727989) Neuregulin-1β (NRG) [Recombinant human neuregulin-1β (NRG1-β1/HRG1-β1 EGF domain), R & D Systems, catalog #396-HB-050] Trastuzumab (Genentech, Inc.) .. EGF/ErbB1 Receptor (D38B1) XP rabbit monoclonal antibody (Cell Signaling, catalog #4267S) ErbB2 Receptor (M45) rabbit polyclonal antibody (Cell Signaling, catalog #3250S) ErbB4 Receptor (111B2) rabbit monoclonal antibody (Cell Signaling, catalog #4795S) AKT (pan; C67E7) rabbit monoclonal antibody (Cell Signaling, catalog #4691S) Phospho-AKT (Ser473) rabbit polyclonal antibody (Cell Signaling, catalog # 9271) p42/44 MAPK (Erk1/2) rabbit polyclonal antibody (Cell Signaling catalog #9102) Phospho- p42/44 MAPK (Erk1/2) (Thr202/Tyr204) rabbit polyclonal (Cell Signaling, catalog #9101) Alpha-tubulin (11H10) rabbit monoclonal antibody (Cell Signaling, catalog #2125) Lamin B1(D4Q4Z) rabbit monoclonal antibody (Cell Signaling, catalog #12586) FOXO3a (D19A7) rabbit monoclonal antibody (Cell Signaling, catalog #12829) Phospho-FOXO3A (S253) rabbit polyclonal antibody (Abcam, catalog #ab47285) CREB (48H2) rabbit monoclonal antibody (Cell Signaling, catalog #9197) Phospho-CREB (Ser133) (87G3) rabbit monoclonal (Cell Signaling, catalog #9198) Amplified Rabbit or Mouse Secondary Antibody Detection Kit (ProteinSimple, catalog #041-126 or 041-127, respectively)

    Article Title: HIV-1 Tat C-mediated regulation of tumor necrosis factor receptor-associated factor-3 by microRNA 32 in human microglia
    Article Snippet: .. Transfection mixtures were prepared in commercial medium (11058–021; Opti-MEM; Invitrogen Corp.), and cells were kept in antibiotic-free medium during transfection. miR-32 overexpression was performed by transfecting the miRNA expression plasmid (SC400329; Origene Technologies, Rockville, MD, USA) using transfection reagent (11668–019; Lipofectamine 2000; Invitrogen Corp.). .. The empty vector was used as vehicle control. miR-32 overexpression was confirmed by qPCR using TaqMan probes specific to miR-32.

    Recombinant:

    Article Title: Use of Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes (hiPSC-CMs) to Monitor Compound Effects on Cardiac Myocyte Signaling Pathways
    Article Snippet: .. Dulbecco's Phosphate-Buffered Saline without Ca2+ and Mg2+ (DPBS, Lonza, Catalog# 17-512F) DMEM, no glucose (Life Technologies, catalog# 11966-025) Sodium pyruvate (Life Technologies, catalog# 11360-070) D-(+)-Galactose (Sigma-Aldrich, catalog# G5388-100G) Paraformaldehyde (Electron Microscopy Science, catalog #15714) Sytox red (Life Technologies, catalog #34859) Goat serum (Rockland, catalog #D204-00-0050) 96-well Acoustic uClear Flat-Bottom Black-Wall Plates (Greiner, catalog #655090) 10X RIPA Lysis Buffer (0.5M Tris-HCl, pH 7.4, 1.5M NaCl, 2.5% deoxycholic acid, 10% NP-40, 10 mM EDTA) (Millipore, catalog #20-188) Sodium Dodecyl Sulfate (SDS, Sigma, catalog #L6026) Protease Inhibitor Cocktail (Sigma, catalog #P8340) Phosphatase Inhibitor Cocktail (Sigma, catalog #P0044) Halt Protease Inhibitor Cocktail, EDTA-free (Thermo Scientific, catalog #87785) M-PER® Mammalian Protein Extraction Reagent (Thermo Scientific, catalog #78501) NE-PER® Nuclear and Cytoplasmic Extraction Reagents (Thermo Scientific, catalog #28833) XDR Charge Separation Master Kit (ProteinSimple, catalog #CBS2001) pI Standard Ladder 3 (ProteinSimple, catalog #040-646) Premix G2, Pharmalyte pH 5-8 Separation Gradient (ProteinSimple, catalog #040-973) DMSO Inhibitor Mix (ProteinSimple, catalog #040-510) Wes™ 12-230 kDa Master Kit (ProteinSimple, catalog #PS-MK01 or PS-MK02) Bicine/CHAPS Lysis Buffer and Sample Diluent (ProteinSimple, catalog #040-764) Pierce BCA Protein Assay Kit (Thermo Scientific, catalog #23225) Lipofectamine RNAiMAX transfection reagent (Life Technologies, catalog #13778) Opti-MEM I Reduced Serum Medium (Life Technologies, catalog #31985-062) ON-TARGETplus SMARTpool Human ErbB2 siRNAs (GE Dharmacon, catalog #L-003126-00-0005) Non-targeting pool control siRNAs (GE Dharmacon, catalog #D-00180-10-05) Lapatinib (National Cancer Institute compound repository, NSC 727989) Neuregulin-1β (NRG) [Recombinant human neuregulin-1β (NRG1-β1/HRG1-β1 EGF domain), R & D Systems, catalog #396-HB-050] Trastuzumab (Genentech, Inc.) .. EGF/ErbB1 Receptor (D38B1) XP rabbit monoclonal antibody (Cell Signaling, catalog #4267S) ErbB2 Receptor (M45) rabbit polyclonal antibody (Cell Signaling, catalog #3250S) ErbB4 Receptor (111B2) rabbit monoclonal antibody (Cell Signaling, catalog #4795S) AKT (pan; C67E7) rabbit monoclonal antibody (Cell Signaling, catalog #4691S) Phospho-AKT (Ser473) rabbit polyclonal antibody (Cell Signaling, catalog # 9271) p42/44 MAPK (Erk1/2) rabbit polyclonal antibody (Cell Signaling catalog #9102) Phospho- p42/44 MAPK (Erk1/2) (Thr202/Tyr204) rabbit polyclonal (Cell Signaling, catalog #9101) Alpha-tubulin (11H10) rabbit monoclonal antibody (Cell Signaling, catalog #2125) Lamin B1(D4Q4Z) rabbit monoclonal antibody (Cell Signaling, catalog #12586) FOXO3a (D19A7) rabbit monoclonal antibody (Cell Signaling, catalog #12829) Phospho-FOXO3A (S253) rabbit polyclonal antibody (Abcam, catalog #ab47285) CREB (48H2) rabbit monoclonal antibody (Cell Signaling, catalog #9197) Phospho-CREB (Ser133) (87G3) rabbit monoclonal (Cell Signaling, catalog #9198) Amplified Rabbit or Mouse Secondary Antibody Detection Kit (ProteinSimple, catalog #041-126 or 041-127, respectively)

    Protease Inhibitor:

    Article Title: Use of Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes (hiPSC-CMs) to Monitor Compound Effects on Cardiac Myocyte Signaling Pathways
    Article Snippet: .. Dulbecco's Phosphate-Buffered Saline without Ca2+ and Mg2+ (DPBS, Lonza, Catalog# 17-512F) DMEM, no glucose (Life Technologies, catalog# 11966-025) Sodium pyruvate (Life Technologies, catalog# 11360-070) D-(+)-Galactose (Sigma-Aldrich, catalog# G5388-100G) Paraformaldehyde (Electron Microscopy Science, catalog #15714) Sytox red (Life Technologies, catalog #34859) Goat serum (Rockland, catalog #D204-00-0050) 96-well Acoustic uClear Flat-Bottom Black-Wall Plates (Greiner, catalog #655090) 10X RIPA Lysis Buffer (0.5M Tris-HCl, pH 7.4, 1.5M NaCl, 2.5% deoxycholic acid, 10% NP-40, 10 mM EDTA) (Millipore, catalog #20-188) Sodium Dodecyl Sulfate (SDS, Sigma, catalog #L6026) Protease Inhibitor Cocktail (Sigma, catalog #P8340) Phosphatase Inhibitor Cocktail (Sigma, catalog #P0044) Halt Protease Inhibitor Cocktail, EDTA-free (Thermo Scientific, catalog #87785) M-PER® Mammalian Protein Extraction Reagent (Thermo Scientific, catalog #78501) NE-PER® Nuclear and Cytoplasmic Extraction Reagents (Thermo Scientific, catalog #28833) XDR Charge Separation Master Kit (ProteinSimple, catalog #CBS2001) pI Standard Ladder 3 (ProteinSimple, catalog #040-646) Premix G2, Pharmalyte pH 5-8 Separation Gradient (ProteinSimple, catalog #040-973) DMSO Inhibitor Mix (ProteinSimple, catalog #040-510) Wes™ 12-230 kDa Master Kit (ProteinSimple, catalog #PS-MK01 or PS-MK02) Bicine/CHAPS Lysis Buffer and Sample Diluent (ProteinSimple, catalog #040-764) Pierce BCA Protein Assay Kit (Thermo Scientific, catalog #23225) Lipofectamine RNAiMAX transfection reagent (Life Technologies, catalog #13778) Opti-MEM I Reduced Serum Medium (Life Technologies, catalog #31985-062) ON-TARGETplus SMARTpool Human ErbB2 siRNAs (GE Dharmacon, catalog #L-003126-00-0005) Non-targeting pool control siRNAs (GE Dharmacon, catalog #D-00180-10-05) Lapatinib (National Cancer Institute compound repository, NSC 727989) Neuregulin-1β (NRG) [Recombinant human neuregulin-1β (NRG1-β1/HRG1-β1 EGF domain), R & D Systems, catalog #396-HB-050] Trastuzumab (Genentech, Inc.) .. EGF/ErbB1 Receptor (D38B1) XP rabbit monoclonal antibody (Cell Signaling, catalog #4267S) ErbB2 Receptor (M45) rabbit polyclonal antibody (Cell Signaling, catalog #3250S) ErbB4 Receptor (111B2) rabbit monoclonal antibody (Cell Signaling, catalog #4795S) AKT (pan; C67E7) rabbit monoclonal antibody (Cell Signaling, catalog #4691S) Phospho-AKT (Ser473) rabbit polyclonal antibody (Cell Signaling, catalog # 9271) p42/44 MAPK (Erk1/2) rabbit polyclonal antibody (Cell Signaling catalog #9102) Phospho- p42/44 MAPK (Erk1/2) (Thr202/Tyr204) rabbit polyclonal (Cell Signaling, catalog #9101) Alpha-tubulin (11H10) rabbit monoclonal antibody (Cell Signaling, catalog #2125) Lamin B1(D4Q4Z) rabbit monoclonal antibody (Cell Signaling, catalog #12586) FOXO3a (D19A7) rabbit monoclonal antibody (Cell Signaling, catalog #12829) Phospho-FOXO3A (S253) rabbit polyclonal antibody (Abcam, catalog #ab47285) CREB (48H2) rabbit monoclonal antibody (Cell Signaling, catalog #9197) Phospho-CREB (Ser133) (87G3) rabbit monoclonal (Cell Signaling, catalog #9198) Amplified Rabbit or Mouse Secondary Antibody Detection Kit (ProteinSimple, catalog #041-126 or 041-127, respectively)

    Infection:

    Article Title: Oncolytic Herpes Virus rRp450 Shows Efficacy in Orthotopic Xenograft Group 3/4 Medulloblastomas and Atypical Teratoid/Rhabdoid Tumors
    Article Snippet: .. Twenty-four hours after seeding, the cells were infected for 2 hr with various MOIs of rRp450 in 0.1 mL of Opti-MEM I reduced serum medium (Thermo Fisher Scientific) at 37°C. .. At the end of the incubation period, the virus was removed, and the cells were maintained in DMEM containing 10% FBS.

    Protein Extraction:

    Article Title: Use of Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes (hiPSC-CMs) to Monitor Compound Effects on Cardiac Myocyte Signaling Pathways
    Article Snippet: .. Dulbecco's Phosphate-Buffered Saline without Ca2+ and Mg2+ (DPBS, Lonza, Catalog# 17-512F) DMEM, no glucose (Life Technologies, catalog# 11966-025) Sodium pyruvate (Life Technologies, catalog# 11360-070) D-(+)-Galactose (Sigma-Aldrich, catalog# G5388-100G) Paraformaldehyde (Electron Microscopy Science, catalog #15714) Sytox red (Life Technologies, catalog #34859) Goat serum (Rockland, catalog #D204-00-0050) 96-well Acoustic uClear Flat-Bottom Black-Wall Plates (Greiner, catalog #655090) 10X RIPA Lysis Buffer (0.5M Tris-HCl, pH 7.4, 1.5M NaCl, 2.5% deoxycholic acid, 10% NP-40, 10 mM EDTA) (Millipore, catalog #20-188) Sodium Dodecyl Sulfate (SDS, Sigma, catalog #L6026) Protease Inhibitor Cocktail (Sigma, catalog #P8340) Phosphatase Inhibitor Cocktail (Sigma, catalog #P0044) Halt Protease Inhibitor Cocktail, EDTA-free (Thermo Scientific, catalog #87785) M-PER® Mammalian Protein Extraction Reagent (Thermo Scientific, catalog #78501) NE-PER® Nuclear and Cytoplasmic Extraction Reagents (Thermo Scientific, catalog #28833) XDR Charge Separation Master Kit (ProteinSimple, catalog #CBS2001) pI Standard Ladder 3 (ProteinSimple, catalog #040-646) Premix G2, Pharmalyte pH 5-8 Separation Gradient (ProteinSimple, catalog #040-973) DMSO Inhibitor Mix (ProteinSimple, catalog #040-510) Wes™ 12-230 kDa Master Kit (ProteinSimple, catalog #PS-MK01 or PS-MK02) Bicine/CHAPS Lysis Buffer and Sample Diluent (ProteinSimple, catalog #040-764) Pierce BCA Protein Assay Kit (Thermo Scientific, catalog #23225) Lipofectamine RNAiMAX transfection reagent (Life Technologies, catalog #13778) Opti-MEM I Reduced Serum Medium (Life Technologies, catalog #31985-062) ON-TARGETplus SMARTpool Human ErbB2 siRNAs (GE Dharmacon, catalog #L-003126-00-0005) Non-targeting pool control siRNAs (GE Dharmacon, catalog #D-00180-10-05) Lapatinib (National Cancer Institute compound repository, NSC 727989) Neuregulin-1β (NRG) [Recombinant human neuregulin-1β (NRG1-β1/HRG1-β1 EGF domain), R & D Systems, catalog #396-HB-050] Trastuzumab (Genentech, Inc.) .. EGF/ErbB1 Receptor (D38B1) XP rabbit monoclonal antibody (Cell Signaling, catalog #4267S) ErbB2 Receptor (M45) rabbit polyclonal antibody (Cell Signaling, catalog #3250S) ErbB4 Receptor (111B2) rabbit monoclonal antibody (Cell Signaling, catalog #4795S) AKT (pan; C67E7) rabbit monoclonal antibody (Cell Signaling, catalog #4691S) Phospho-AKT (Ser473) rabbit polyclonal antibody (Cell Signaling, catalog # 9271) p42/44 MAPK (Erk1/2) rabbit polyclonal antibody (Cell Signaling catalog #9102) Phospho- p42/44 MAPK (Erk1/2) (Thr202/Tyr204) rabbit polyclonal (Cell Signaling, catalog #9101) Alpha-tubulin (11H10) rabbit monoclonal antibody (Cell Signaling, catalog #2125) Lamin B1(D4Q4Z) rabbit monoclonal antibody (Cell Signaling, catalog #12586) FOXO3a (D19A7) rabbit monoclonal antibody (Cell Signaling, catalog #12829) Phospho-FOXO3A (S253) rabbit polyclonal antibody (Abcam, catalog #ab47285) CREB (48H2) rabbit monoclonal antibody (Cell Signaling, catalog #9197) Phospho-CREB (Ser133) (87G3) rabbit monoclonal (Cell Signaling, catalog #9198) Amplified Rabbit or Mouse Secondary Antibody Detection Kit (ProteinSimple, catalog #041-126 or 041-127, respectively)

    other:

    Article Title: Secreted dual reporter assay with Gaussia luciferase and the red fluorescent protein mCherry
    Article Snippet: Reagents Native coelenterazine was from BIOSYNTH, "Opti-MEM I Reduced Serum Medium, without Phenol Red" (Opti-MEM) from Life Technologies.

    Cell Culture:

    Article Title: IGFBP7 induces apoptosis of acute myeloid leukemia cells and synergizes with chemotherapy in suppression of leukemia cell survival
    Article Snippet: .. Cells cultured under low-serum conditions were either cultured in RPMI-1640 with 1% FCS or in Opti-Mem Reduced-Serum Medium (used in the experiments illustrated in , and ; Life Technologies, Bleiswijk, The Netherlands) with 1% FCS. .. IGFBP7-overexpressing AML cell lines were generated by lentiviral transduction with pCDH-CMV-EF-1-puro-IGFBP7 (System Biosciences).

    Expressing:

    Article Title: HIV-1 Tat C-mediated regulation of tumor necrosis factor receptor-associated factor-3 by microRNA 32 in human microglia
    Article Snippet: .. Transfection mixtures were prepared in commercial medium (11058–021; Opti-MEM; Invitrogen Corp.), and cells were kept in antibiotic-free medium during transfection. miR-32 overexpression was performed by transfecting the miRNA expression plasmid (SC400329; Origene Technologies, Rockville, MD, USA) using transfection reagent (11668–019; Lipofectamine 2000; Invitrogen Corp.). .. The empty vector was used as vehicle control. miR-32 overexpression was confirmed by qPCR using TaqMan probes specific to miR-32.

    BIA-KA:

    Article Title: Use of Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes (hiPSC-CMs) to Monitor Compound Effects on Cardiac Myocyte Signaling Pathways
    Article Snippet: .. Dulbecco's Phosphate-Buffered Saline without Ca2+ and Mg2+ (DPBS, Lonza, Catalog# 17-512F) DMEM, no glucose (Life Technologies, catalog# 11966-025) Sodium pyruvate (Life Technologies, catalog# 11360-070) D-(+)-Galactose (Sigma-Aldrich, catalog# G5388-100G) Paraformaldehyde (Electron Microscopy Science, catalog #15714) Sytox red (Life Technologies, catalog #34859) Goat serum (Rockland, catalog #D204-00-0050) 96-well Acoustic uClear Flat-Bottom Black-Wall Plates (Greiner, catalog #655090) 10X RIPA Lysis Buffer (0.5M Tris-HCl, pH 7.4, 1.5M NaCl, 2.5% deoxycholic acid, 10% NP-40, 10 mM EDTA) (Millipore, catalog #20-188) Sodium Dodecyl Sulfate (SDS, Sigma, catalog #L6026) Protease Inhibitor Cocktail (Sigma, catalog #P8340) Phosphatase Inhibitor Cocktail (Sigma, catalog #P0044) Halt Protease Inhibitor Cocktail, EDTA-free (Thermo Scientific, catalog #87785) M-PER® Mammalian Protein Extraction Reagent (Thermo Scientific, catalog #78501) NE-PER® Nuclear and Cytoplasmic Extraction Reagents (Thermo Scientific, catalog #28833) XDR Charge Separation Master Kit (ProteinSimple, catalog #CBS2001) pI Standard Ladder 3 (ProteinSimple, catalog #040-646) Premix G2, Pharmalyte pH 5-8 Separation Gradient (ProteinSimple, catalog #040-973) DMSO Inhibitor Mix (ProteinSimple, catalog #040-510) Wes™ 12-230 kDa Master Kit (ProteinSimple, catalog #PS-MK01 or PS-MK02) Bicine/CHAPS Lysis Buffer and Sample Diluent (ProteinSimple, catalog #040-764) Pierce BCA Protein Assay Kit (Thermo Scientific, catalog #23225) Lipofectamine RNAiMAX transfection reagent (Life Technologies, catalog #13778) Opti-MEM I Reduced Serum Medium (Life Technologies, catalog #31985-062) ON-TARGETplus SMARTpool Human ErbB2 siRNAs (GE Dharmacon, catalog #L-003126-00-0005) Non-targeting pool control siRNAs (GE Dharmacon, catalog #D-00180-10-05) Lapatinib (National Cancer Institute compound repository, NSC 727989) Neuregulin-1β (NRG) [Recombinant human neuregulin-1β (NRG1-β1/HRG1-β1 EGF domain), R & D Systems, catalog #396-HB-050] Trastuzumab (Genentech, Inc.) .. EGF/ErbB1 Receptor (D38B1) XP rabbit monoclonal antibody (Cell Signaling, catalog #4267S) ErbB2 Receptor (M45) rabbit polyclonal antibody (Cell Signaling, catalog #3250S) ErbB4 Receptor (111B2) rabbit monoclonal antibody (Cell Signaling, catalog #4795S) AKT (pan; C67E7) rabbit monoclonal antibody (Cell Signaling, catalog #4691S) Phospho-AKT (Ser473) rabbit polyclonal antibody (Cell Signaling, catalog # 9271) p42/44 MAPK (Erk1/2) rabbit polyclonal antibody (Cell Signaling catalog #9102) Phospho- p42/44 MAPK (Erk1/2) (Thr202/Tyr204) rabbit polyclonal (Cell Signaling, catalog #9101) Alpha-tubulin (11H10) rabbit monoclonal antibody (Cell Signaling, catalog #2125) Lamin B1(D4Q4Z) rabbit monoclonal antibody (Cell Signaling, catalog #12586) FOXO3a (D19A7) rabbit monoclonal antibody (Cell Signaling, catalog #12829) Phospho-FOXO3A (S253) rabbit polyclonal antibody (Abcam, catalog #ab47285) CREB (48H2) rabbit monoclonal antibody (Cell Signaling, catalog #9197) Phospho-CREB (Ser133) (87G3) rabbit monoclonal (Cell Signaling, catalog #9198) Amplified Rabbit or Mouse Secondary Antibody Detection Kit (ProteinSimple, catalog #041-126 or 041-127, respectively)

    Modification:

    Article Title: Formulation of pH responsive peptides as inhalable dry powders for pulmonary delivery of nucleic acids
    Article Snippet: .. Dulbecco’s modified eagle medium (DMEM), OptiMEM-1 reduced serum medium, antibiotic-animycotic liquid, fetal bovine serum (FBS) and Lipofectamine™ 2000 were purchased from Invitrogen (CA, USA). .. The luciferase assay system was purchased from Promega (Madison, WI, USA).

    Lysis:

    Article Title: Use of Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes (hiPSC-CMs) to Monitor Compound Effects on Cardiac Myocyte Signaling Pathways
    Article Snippet: .. Dulbecco's Phosphate-Buffered Saline without Ca2+ and Mg2+ (DPBS, Lonza, Catalog# 17-512F) DMEM, no glucose (Life Technologies, catalog# 11966-025) Sodium pyruvate (Life Technologies, catalog# 11360-070) D-(+)-Galactose (Sigma-Aldrich, catalog# G5388-100G) Paraformaldehyde (Electron Microscopy Science, catalog #15714) Sytox red (Life Technologies, catalog #34859) Goat serum (Rockland, catalog #D204-00-0050) 96-well Acoustic uClear Flat-Bottom Black-Wall Plates (Greiner, catalog #655090) 10X RIPA Lysis Buffer (0.5M Tris-HCl, pH 7.4, 1.5M NaCl, 2.5% deoxycholic acid, 10% NP-40, 10 mM EDTA) (Millipore, catalog #20-188) Sodium Dodecyl Sulfate (SDS, Sigma, catalog #L6026) Protease Inhibitor Cocktail (Sigma, catalog #P8340) Phosphatase Inhibitor Cocktail (Sigma, catalog #P0044) Halt Protease Inhibitor Cocktail, EDTA-free (Thermo Scientific, catalog #87785) M-PER® Mammalian Protein Extraction Reagent (Thermo Scientific, catalog #78501) NE-PER® Nuclear and Cytoplasmic Extraction Reagents (Thermo Scientific, catalog #28833) XDR Charge Separation Master Kit (ProteinSimple, catalog #CBS2001) pI Standard Ladder 3 (ProteinSimple, catalog #040-646) Premix G2, Pharmalyte pH 5-8 Separation Gradient (ProteinSimple, catalog #040-973) DMSO Inhibitor Mix (ProteinSimple, catalog #040-510) Wes™ 12-230 kDa Master Kit (ProteinSimple, catalog #PS-MK01 or PS-MK02) Bicine/CHAPS Lysis Buffer and Sample Diluent (ProteinSimple, catalog #040-764) Pierce BCA Protein Assay Kit (Thermo Scientific, catalog #23225) Lipofectamine RNAiMAX transfection reagent (Life Technologies, catalog #13778) Opti-MEM I Reduced Serum Medium (Life Technologies, catalog #31985-062) ON-TARGETplus SMARTpool Human ErbB2 siRNAs (GE Dharmacon, catalog #L-003126-00-0005) Non-targeting pool control siRNAs (GE Dharmacon, catalog #D-00180-10-05) Lapatinib (National Cancer Institute compound repository, NSC 727989) Neuregulin-1β (NRG) [Recombinant human neuregulin-1β (NRG1-β1/HRG1-β1 EGF domain), R & D Systems, catalog #396-HB-050] Trastuzumab (Genentech, Inc.) .. EGF/ErbB1 Receptor (D38B1) XP rabbit monoclonal antibody (Cell Signaling, catalog #4267S) ErbB2 Receptor (M45) rabbit polyclonal antibody (Cell Signaling, catalog #3250S) ErbB4 Receptor (111B2) rabbit monoclonal antibody (Cell Signaling, catalog #4795S) AKT (pan; C67E7) rabbit monoclonal antibody (Cell Signaling, catalog #4691S) Phospho-AKT (Ser473) rabbit polyclonal antibody (Cell Signaling, catalog # 9271) p42/44 MAPK (Erk1/2) rabbit polyclonal antibody (Cell Signaling catalog #9102) Phospho- p42/44 MAPK (Erk1/2) (Thr202/Tyr204) rabbit polyclonal (Cell Signaling, catalog #9101) Alpha-tubulin (11H10) rabbit monoclonal antibody (Cell Signaling, catalog #2125) Lamin B1(D4Q4Z) rabbit monoclonal antibody (Cell Signaling, catalog #12586) FOXO3a (D19A7) rabbit monoclonal antibody (Cell Signaling, catalog #12829) Phospho-FOXO3A (S253) rabbit polyclonal antibody (Abcam, catalog #ab47285) CREB (48H2) rabbit monoclonal antibody (Cell Signaling, catalog #9197) Phospho-CREB (Ser133) (87G3) rabbit monoclonal (Cell Signaling, catalog #9198) Amplified Rabbit or Mouse Secondary Antibody Detection Kit (ProteinSimple, catalog #041-126 or 041-127, respectively)

    Over Expression:

    Article Title: HIV-1 Tat C-mediated regulation of tumor necrosis factor receptor-associated factor-3 by microRNA 32 in human microglia
    Article Snippet: .. Transfection mixtures were prepared in commercial medium (11058–021; Opti-MEM; Invitrogen Corp.), and cells were kept in antibiotic-free medium during transfection. miR-32 overexpression was performed by transfecting the miRNA expression plasmid (SC400329; Origene Technologies, Rockville, MD, USA) using transfection reagent (11668–019; Lipofectamine 2000; Invitrogen Corp.). .. The empty vector was used as vehicle control. miR-32 overexpression was confirmed by qPCR using TaqMan probes specific to miR-32.

    Plasmid Preparation:

    Article Title: HIV-1 Tat C-mediated regulation of tumor necrosis factor receptor-associated factor-3 by microRNA 32 in human microglia
    Article Snippet: .. Transfection mixtures were prepared in commercial medium (11058–021; Opti-MEM; Invitrogen Corp.), and cells were kept in antibiotic-free medium during transfection. miR-32 overexpression was performed by transfecting the miRNA expression plasmid (SC400329; Origene Technologies, Rockville, MD, USA) using transfection reagent (11668–019; Lipofectamine 2000; Invitrogen Corp.). .. The empty vector was used as vehicle control. miR-32 overexpression was confirmed by qPCR using TaqMan probes specific to miR-32.

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    Thermo Fisher opti mem reduced serum medium
    Opti Mem Reduced Serum Medium, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 715 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/opti mem reduced serum medium/product/Thermo Fisher
    Average 92 stars, based on 715 article reviews
    Price from $9.99 to $1999.99
    opti mem reduced serum medium - by Bioz Stars, 2020-07
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