Structured Review

Millipore l lysine
Leader bleb-based migration is resistant to Src inhibition. a Ventral Z-section of an EGFP-RLC- and FusionRed-F-tractin-expressing A375 cell treated with 500 nM Dasatinib 90 min prior to being highly confined. b Quantitative evaluation of ( a ) for the percent migratory (black bars; leader bleb-based) cells with ( n = 15) or without ( n = 38) Dasatinib treatment. c Normalized cortical F-actin density in A375 cells with ( n = 12) or without ( n = 11) Dasatinib treatment as determined by phalloidin staining of the actin cortex. Fluorescence intensity measurements were measured from the central Z-section of cells freshly plated on <t>poly-L-lysine-coated</t> glass. d , e Cortex tension and intracellular pressure of A375 cells with ( n = 27) or without ( n = 28) Dasatinib treatment. Measurements were taken on freshly plated, round, A375 cells on uncoated glass. Intracellular pressures were calculated from cortical tension and cell radii measured by light microscopy. f Central Z-section of a YFP-tagged tandem SH2 domain construct (YFP-dSH2) expressed in a highly confined A375 cell. g Ventral Z-section of a YFP-dSH2 expressing A375 cell on <t>fibronectin-coated</t> glass. All data are representative of at least three independent experiments. Statistical significance was determined by one-way ANOVA. *** p ≤ 0.001
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Images

1) Product Images from "c-Src activity is differentially required by cancer cell motility modes"

Article Title: c-Src activity is differentially required by cancer cell motility modes

Journal: Oncogene

doi: 10.1038/s41388-017-0071-5

Leader bleb-based migration is resistant to Src inhibition. a Ventral Z-section of an EGFP-RLC- and FusionRed-F-tractin-expressing A375 cell treated with 500 nM Dasatinib 90 min prior to being highly confined. b Quantitative evaluation of ( a ) for the percent migratory (black bars; leader bleb-based) cells with ( n = 15) or without ( n = 38) Dasatinib treatment. c Normalized cortical F-actin density in A375 cells with ( n = 12) or without ( n = 11) Dasatinib treatment as determined by phalloidin staining of the actin cortex. Fluorescence intensity measurements were measured from the central Z-section of cells freshly plated on poly-L-lysine-coated glass. d , e Cortex tension and intracellular pressure of A375 cells with ( n = 27) or without ( n = 28) Dasatinib treatment. Measurements were taken on freshly plated, round, A375 cells on uncoated glass. Intracellular pressures were calculated from cortical tension and cell radii measured by light microscopy. f Central Z-section of a YFP-tagged tandem SH2 domain construct (YFP-dSH2) expressed in a highly confined A375 cell. g Ventral Z-section of a YFP-dSH2 expressing A375 cell on fibronectin-coated glass. All data are representative of at least three independent experiments. Statistical significance was determined by one-way ANOVA. *** p ≤ 0.001
Figure Legend Snippet: Leader bleb-based migration is resistant to Src inhibition. a Ventral Z-section of an EGFP-RLC- and FusionRed-F-tractin-expressing A375 cell treated with 500 nM Dasatinib 90 min prior to being highly confined. b Quantitative evaluation of ( a ) for the percent migratory (black bars; leader bleb-based) cells with ( n = 15) or without ( n = 38) Dasatinib treatment. c Normalized cortical F-actin density in A375 cells with ( n = 12) or without ( n = 11) Dasatinib treatment as determined by phalloidin staining of the actin cortex. Fluorescence intensity measurements were measured from the central Z-section of cells freshly plated on poly-L-lysine-coated glass. d , e Cortex tension and intracellular pressure of A375 cells with ( n = 27) or without ( n = 28) Dasatinib treatment. Measurements were taken on freshly plated, round, A375 cells on uncoated glass. Intracellular pressures were calculated from cortical tension and cell radii measured by light microscopy. f Central Z-section of a YFP-tagged tandem SH2 domain construct (YFP-dSH2) expressed in a highly confined A375 cell. g Ventral Z-section of a YFP-dSH2 expressing A375 cell on fibronectin-coated glass. All data are representative of at least three independent experiments. Statistical significance was determined by one-way ANOVA. *** p ≤ 0.001

Techniques Used: Migration, Inhibition, Expressing, Staining, Fluorescence, Light Microscopy, Construct

2) Product Images from "Cell adhesion to the extracellular matrix protein fibronectin modulates radiation-dependent G2 phase arrest involving integrin-linked kinase (ILK) and glycogen synthase kinase-3β (GSK-3β) in vitro"

Article Title: Cell adhesion to the extracellular matrix protein fibronectin modulates radiation-dependent G2 phase arrest involving integrin-linked kinase (ILK) and glycogen synthase kinase-3β (GSK-3β) in vitro

Journal: British Journal of Cancer

doi: 10.1038/sj.bjc.6600912

Analysis of substrate-dependent clonogenic cell survival was accomplished by plating human lung carcinoma cells A549 and normal human lung fibroblasts CCD32 24 h prior to irradiation onto polystyrene (•) or a specific protein (▴: FN, LA, BSA, poly- L ). P
Figure Legend Snippet: Analysis of substrate-dependent clonogenic cell survival was accomplished by plating human lung carcinoma cells A549 and normal human lung fibroblasts CCD32 24 h prior to irradiation onto polystyrene (•) or a specific protein (▴: FN, LA, BSA, poly- L ). P

Techniques Used: Irradiation

3) Product Images from "c-Src activity is differentially required by cancer cell motility modes"

Article Title: c-Src activity is differentially required by cancer cell motility modes

Journal: Oncogene

doi: 10.1038/s41388-017-0071-5

Leader bleb-based migration is resistant to Src inhibition. a Ventral Z-section of an EGFP-RLC- and FusionRed-F-tractin-expressing A375 cell treated with 500 nM Dasatinib 90 min prior to being highly confined. b Quantitative evaluation of ( a ) for the percent migratory (black bars; leader bleb-based) cells with ( n = 15) or without ( n = 38) Dasatinib treatment. c Normalized cortical F-actin density in A375 cells with ( n = 12) or without ( n = 11) Dasatinib treatment as determined by phalloidin staining of the actin cortex. Fluorescence intensity measurements were measured from the central Z-section of cells freshly plated on poly-L-lysine-coated glass. d , e Cortex tension and intracellular pressure of A375 cells with ( n = 27) or without ( n = 28) Dasatinib treatment. Measurements were taken on freshly plated, round, A375 cells on uncoated glass. Intracellular pressures were calculated from cortical tension and cell radii measured by light microscopy. f Central Z-section of a YFP-tagged tandem SH2 domain construct (YFP-dSH2) expressed in a highly confined A375 cell. g Ventral Z-section of a YFP-dSH2 expressing A375 cell on fibronectin-coated glass. All data are representative of at least three independent experiments. Statistical significance was determined by one-way ANOVA. *** p ≤ 0.001
Figure Legend Snippet: Leader bleb-based migration is resistant to Src inhibition. a Ventral Z-section of an EGFP-RLC- and FusionRed-F-tractin-expressing A375 cell treated with 500 nM Dasatinib 90 min prior to being highly confined. b Quantitative evaluation of ( a ) for the percent migratory (black bars; leader bleb-based) cells with ( n = 15) or without ( n = 38) Dasatinib treatment. c Normalized cortical F-actin density in A375 cells with ( n = 12) or without ( n = 11) Dasatinib treatment as determined by phalloidin staining of the actin cortex. Fluorescence intensity measurements were measured from the central Z-section of cells freshly plated on poly-L-lysine-coated glass. d , e Cortex tension and intracellular pressure of A375 cells with ( n = 27) or without ( n = 28) Dasatinib treatment. Measurements were taken on freshly plated, round, A375 cells on uncoated glass. Intracellular pressures were calculated from cortical tension and cell radii measured by light microscopy. f Central Z-section of a YFP-tagged tandem SH2 domain construct (YFP-dSH2) expressed in a highly confined A375 cell. g Ventral Z-section of a YFP-dSH2 expressing A375 cell on fibronectin-coated glass. All data are representative of at least three independent experiments. Statistical significance was determined by one-way ANOVA. *** p ≤ 0.001

Techniques Used: Migration, Inhibition, Expressing, Staining, Fluorescence, Light Microscopy, Construct

4) Product Images from "Role of TC21/R-Ras2 in enhanced migration of neurofibromin-deficient Schwann cells"

Article Title: Role of TC21/R-Ras2 in enhanced migration of neurofibromin-deficient Schwann cells

Journal: Oncogene

doi: 10.1038/sj.onc.1207075

Nf1 -deficient MSCs exhibit increased migration in vitro . Passage-matched Nf1 +/+, Nf1 +/−, and Nf1 −/− MSCs were plated onto poly-L-lysine-coated transwells and counted 16 h later. ( a ) N2 medium (N2) was added to the lower chamber of the transwell. ( b ) Conditioned medium (CM) derived from Nf1 +/+, Nf1 +/−, or Nf1 −/− MSCs or N2 was added to the lower chamber of the transwell. Migration was quantified by counting the number of cells present on the entire transwell filter ( a ) or in four fields ( b ) using a fluorescent microscope. The average of duplicate samples is shown; duplicates varied by less than 10%. The number of cells that migrated was normalized to the total number of cells plated. ** P
Figure Legend Snippet: Nf1 -deficient MSCs exhibit increased migration in vitro . Passage-matched Nf1 +/+, Nf1 +/−, and Nf1 −/− MSCs were plated onto poly-L-lysine-coated transwells and counted 16 h later. ( a ) N2 medium (N2) was added to the lower chamber of the transwell. ( b ) Conditioned medium (CM) derived from Nf1 +/+, Nf1 +/−, or Nf1 −/− MSCs or N2 was added to the lower chamber of the transwell. Migration was quantified by counting the number of cells present on the entire transwell filter ( a ) or in four fields ( b ) using a fluorescent microscope. The average of duplicate samples is shown; duplicates varied by less than 10%. The number of cells that migrated was normalized to the total number of cells plated. ** P

Techniques Used: Migration, In Vitro, Derivative Assay, Microscopy

Related Articles

Cytometry:

Article Title: Dynamics of genome reorganization during human cardiogenesis reveal an RBM20-dependent splicing factory
Article Snippet: This well-established procedure efficiently removed contaminating non-CM cells (which do not tolerate lactate metabolic selection), thus highly enriching for CM ( > 95% as assessed by Flow Cytometry for cTnT and immunofluorescence for α-actinin). .. Poly-Lysine slides (Sigma-Aldrich, P0425) were pre-coated with 0.5 or 2 μg mL−1 rhLaminin521 (for hESCs or CMs, respectively) dispersed on a ~2 cm2 spot outlined by a hydrophobic pen for 2 h at 37 °C.

Co-Culture Assay:

Article Title: Engineering a haematopoietic stem cell niche by revitalizing mesenchymal stromal cells
Article Snippet: .. After co-culture in serum-free medium supplemented with SCF (20ng/ml) and TPO (10ng/ml) for 6 days, Lin− Sca-1+ c-Kit+ CD48− CD150+ HSCs were directly sorted on polylysine-coated slides (P0425–72AE; Sigma), 500–2000 cells per slide, incubated for 10 min, fixed with 4 % PFA for 10 min at room temperature, and permeabilised with 0.2 % Triton X-100 for 20 min at room temperature. ..

Incubation:

Article Title: Adrenergic nerve degeneration in bone marrow drives aging of the hematopoietic stem cell niche
Article Snippet: .. Lineage− CD48− Sca-1+ c-Kit+ CD150+ HSCs were directly sorted on poly-lysine coated slides (P0425-72AE; Sigma), 500–2000 cells per slide, incubated for 10 min, fixed with 4% PFA for 10 min at room temperature, and permeabilized with 0.2% Triton X-100 for 20 min at room temperature. ..

Article Title: Engineering a haematopoietic stem cell niche by revitalizing mesenchymal stromal cells
Article Snippet: .. After co-culture in serum-free medium supplemented with SCF (20ng/ml) and TPO (10ng/ml) for 6 days, Lin− Sca-1+ c-Kit+ CD48− CD150+ HSCs were directly sorted on polylysine-coated slides (P0425–72AE; Sigma), 500–2000 cells per slide, incubated for 10 min, fixed with 4 % PFA for 10 min at room temperature, and permeabilised with 0.2 % Triton X-100 for 20 min at room temperature. ..

Article Title: Zinc finger protein Zfp335 is required for the formation of the naïve T cell compartment
Article Snippet: Immunofluorescence Mature CD4SP thymocytes were sorted and allowed to adhere to poly-L-lysine-coated glass slides (P0425; Sigma–Aldrich, St. Louis, MO). .. For Zfp335 detection, slides were incubated at RT with primary antibody (A300-797A, A300-798A; Bethyl Laboratories, Montgomery, TX) for at least 3 hr, followed by biotin goat anti-rabbit (BD Biosciences) and lastly SA-Cy3 (Jackson Immunoresearch, West Grove, PA).

Article Title: Necessity and Contingency in Developmental Genetic Screens: EGF, Wnt, and Semaphorin Pathways in Vulval Induction of the Nematode Oscheius tipulae
Article Snippet: In brief, OP50 -grown populations were washed three times in distilled water and placed onto poly-l -lysine-coated (P0425-72EA; Sigma [Sigma Chemical], St. Louis, MO) slides prior to freeze-cracking. .. Worms with an open cuticle were incubated in antibody buffer with the mouse MH27 antibody against the epithelial cell adherent junctions ( ).

Transfection:

Article Title: Novel initiator caspase reporters uncover previously unknown features of caspase-activating cells
Article Snippet: Paragraph title: Cell culture and transfection of S2 and S2-R+ cells ... In most experiments, cells were grown on normal coverslips placed in 6-well plates; however, coverslips coated with poly-lysine (Sigma, P0425-72EA) were used to facilitate the visualisation of cell projections and active actin in Fig. S1E .

Expressing:

Article Title: Study of amyloids using yeast
Article Snippet: Yeast cells expressing fluorescent proteins can be observed using a standard fluorescent microscope with the appropriate filter sets using 60x or 100x objective lenses. .. To further reduce movement microscope slides can be coated with poly l-lysine (Sigma P0425) or the lectin concanavalin A, which both bind the yeast cell wall.

Translocation Assay:

Article Title: MC-PPEA as a new and more potent inhibitor of CLP-induced sepsis and pulmonary inflammation than FK866
Article Snippet: .. Immunofluorescence imaging of nuclear translocation of NFκB in A549 cells A549 cells were seeded on poly-L-lysine-coated glass slides (Catalog#: p0425; Sigma-Aldrich) and cultured in a 35 mm Petri dish with 3 mL of media at 37°C in a humidified atmosphere of 5% CO2 . ..

Flow Cytometry:

Article Title: Dynamics of genome reorganization during human cardiogenesis reveal an RBM20-dependent splicing factory
Article Snippet: This well-established procedure efficiently removed contaminating non-CM cells (which do not tolerate lactate metabolic selection), thus highly enriching for CM ( > 95% as assessed by Flow Cytometry for cTnT and immunofluorescence for α-actinin). .. Poly-Lysine slides (Sigma-Aldrich, P0425) were pre-coated with 0.5 or 2 μg mL−1 rhLaminin521 (for hESCs or CMs, respectively) dispersed on a ~2 cm2 spot outlined by a hydrophobic pen for 2 h at 37 °C.

Transferring:

Article Title: Study of amyloids using yeast
Article Snippet: Slightly tapping the cover slip with a pipette tip after it has been applied but before the slide is placed on the microscope also reduces movement of the cells. .. To further reduce movement microscope slides can be coated with poly l-lysine (Sigma P0425) or the lectin concanavalin A, which both bind the yeast cell wall.

Cell Culture:

Article Title: Dynamics of genome reorganization during human cardiogenesis reveal an RBM20-dependent splicing factory
Article Snippet: CM were then cultured in RPMI-B27 for 4 more days before being collected for 3D DNA FISH. .. Poly-Lysine slides (Sigma-Aldrich, P0425) were pre-coated with 0.5 or 2 μg mL−1 rhLaminin521 (for hESCs or CMs, respectively) dispersed on a ~2 cm2 spot outlined by a hydrophobic pen for 2 h at 37 °C.

Article Title: Novel initiator caspase reporters uncover previously unknown features of caspase-activating cells
Article Snippet: Paragraph title: Cell culture and transfection of S2 and S2-R+ cells ... In most experiments, cells were grown on normal coverslips placed in 6-well plates; however, coverslips coated with poly-lysine (Sigma, P0425-72EA) were used to facilitate the visualisation of cell projections and active actin in Fig. S1E .

Article Title: MC-PPEA as a new and more potent inhibitor of CLP-induced sepsis and pulmonary inflammation than FK866
Article Snippet: .. Immunofluorescence imaging of nuclear translocation of NFκB in A549 cells A549 cells were seeded on poly-L-lysine-coated glass slides (Catalog#: p0425; Sigma-Aldrich) and cultured in a 35 mm Petri dish with 3 mL of media at 37°C in a humidified atmosphere of 5% CO2 . ..

Labeling:

Article Title: Necessity and Contingency in Developmental Genetic Screens: EGF, Wnt, and Semaphorin Pathways in Vulval Induction of the Nematode Oscheius tipulae
Article Snippet: In brief, OP50 -grown populations were washed three times in distilled water and placed onto poly-l -lysine-coated (P0425-72EA; Sigma [Sigma Chemical], St. Louis, MO) slides prior to freeze-cracking. .. As secondary antibody, we used the goat anti-mouse antibody from Abcam labeled fluorescently with Alexa Fluor 488 (reference #ab150113).

Article Title: The QTL GNP1 Encodes GA20ox1, Which Increases Grain Number and Yield by Increasing Cytokinin Activity in Rice Panicle Meristems
Article Snippet: The samples were then embedded in Paraplast Plus (Sigma, P3683), cut into 10 μm sections and mounted on pre-coated poly-prep slides (Sigma, P0425). .. Digoxigenin-labeled RNA probes were prepared following the instructions of the DIG RNA labeling kit (SP6/T7) (Roche, 11175025910).

Inhibition:

Article Title: Dynamics of genome reorganization during human cardiogenesis reveal an RBM20-dependent splicing factory
Article Snippet: Poly-Lysine slides (Sigma-Aldrich, P0425) were pre-coated with 0.5 or 2 μg mL−1 rhLaminin521 (for hESCs or CMs, respectively) dispersed on a ~2 cm2 spot outlined by a hydrophobic pen for 2 h at 37 °C. .. For transcription inhibition experiments, 5 µM Actinomycin D was added to the culture media both 3 h before collecting the cells and during the seeding procedure.

Imaging:

Article Title: Adrenergic nerve degeneration in bone marrow drives aging of the hematopoietic stem cell niche
Article Snippet: Paragraph title: Immunofluorescence staining and imaging of sorted HSCs ... Lineage− CD48− Sca-1+ c-Kit+ CD150+ HSCs were directly sorted on poly-lysine coated slides (P0425-72AE; Sigma), 500–2000 cells per slide, incubated for 10 min, fixed with 4% PFA for 10 min at room temperature, and permeabilized with 0.2% Triton X-100 for 20 min at room temperature.

Article Title: Engineering a haematopoietic stem cell niche by revitalizing mesenchymal stromal cells
Article Snippet: Paragraph title: Immunofluorescence staining and imaging of sorted HSCs ... After co-culture in serum-free medium supplemented with SCF (20ng/ml) and TPO (10ng/ml) for 6 days, Lin− Sca-1+ c-Kit+ CD48− CD150+ HSCs were directly sorted on polylysine-coated slides (P0425–72AE; Sigma), 500–2000 cells per slide, incubated for 10 min, fixed with 4 % PFA for 10 min at room temperature, and permeabilised with 0.2 % Triton X-100 for 20 min at room temperature.

Article Title: MC-PPEA as a new and more potent inhibitor of CLP-induced sepsis and pulmonary inflammation than FK866
Article Snippet: .. Immunofluorescence imaging of nuclear translocation of NFκB in A549 cells A549 cells were seeded on poly-L-lysine-coated glass slides (Catalog#: p0425; Sigma-Aldrich) and cultured in a 35 mm Petri dish with 3 mL of media at 37°C in a humidified atmosphere of 5% CO2 . ..

Immunostaining:

Article Title: Aurora-A mediated histone H3 phosphorylation of threonine 118 controls condensin I and cohesin occupancy in mitosis
Article Snippet: .. C. elegans immunostaining Embryos from adult hermaphrodites were picked into 10 μl egg buffer on a Poly-L-Lysine coated glass slide (Sigma, St Louis, MO P0425). ..

Article Title: Necessity and Contingency in Developmental Genetic Screens: EGF, Wnt, and Semaphorin Pathways in Vulval Induction of the Nematode Oscheius tipulae
Article Snippet: Immunofluorescence staining Bleach-synchronized larvae and mixed-stage populations were fixed, and permeabilized for immunostaining using previously described methods ( ; , ). .. In brief, OP50 -grown populations were washed three times in distilled water and placed onto poly-l -lysine-coated (P0425-72EA; Sigma [Sigma Chemical], St. Louis, MO) slides prior to freeze-cracking.

Immunofluorescence:

Article Title: Dynamics of genome reorganization during human cardiogenesis reveal an RBM20-dependent splicing factory
Article Snippet: This well-established procedure efficiently removed contaminating non-CM cells (which do not tolerate lactate metabolic selection), thus highly enriching for CM ( > 95% as assessed by Flow Cytometry for cTnT and immunofluorescence for α-actinin). .. Poly-Lysine slides (Sigma-Aldrich, P0425) were pre-coated with 0.5 or 2 μg mL−1 rhLaminin521 (for hESCs or CMs, respectively) dispersed on a ~2 cm2 spot outlined by a hydrophobic pen for 2 h at 37 °C.

Article Title: Adrenergic nerve degeneration in bone marrow drives aging of the hematopoietic stem cell niche
Article Snippet: Paragraph title: Immunofluorescence staining and imaging of sorted HSCs ... Lineage− CD48− Sca-1+ c-Kit+ CD150+ HSCs were directly sorted on poly-lysine coated slides (P0425-72AE; Sigma), 500–2000 cells per slide, incubated for 10 min, fixed with 4% PFA for 10 min at room temperature, and permeabilized with 0.2% Triton X-100 for 20 min at room temperature.

Article Title: Engineering a haematopoietic stem cell niche by revitalizing mesenchymal stromal cells
Article Snippet: Paragraph title: Immunofluorescence staining and imaging of sorted HSCs ... After co-culture in serum-free medium supplemented with SCF (20ng/ml) and TPO (10ng/ml) for 6 days, Lin− Sca-1+ c-Kit+ CD48− CD150+ HSCs were directly sorted on polylysine-coated slides (P0425–72AE; Sigma), 500–2000 cells per slide, incubated for 10 min, fixed with 4 % PFA for 10 min at room temperature, and permeabilised with 0.2 % Triton X-100 for 20 min at room temperature.

Article Title: Myeloid progenitor cluster formation drives emergency and leukemic myelopoiesis
Article Snippet: .. Immunofluorescence on purified cells GMP (2,000 cells/slide) were directly sorted onto poly-lysine coated slides (Sigma-Aldrich, P0425-72EA) and let settle down for 15 min at RT. .. Cells were washed 3x for 5 min with PBS at RT between each staining step.

Article Title: MC-PPEA as a new and more potent inhibitor of CLP-induced sepsis and pulmonary inflammation than FK866
Article Snippet: .. Immunofluorescence imaging of nuclear translocation of NFκB in A549 cells A549 cells were seeded on poly-L-lysine-coated glass slides (Catalog#: p0425; Sigma-Aldrich) and cultured in a 35 mm Petri dish with 3 mL of media at 37°C in a humidified atmosphere of 5% CO2 . ..

Article Title: Zinc finger protein Zfp335 is required for the formation of the naïve T cell compartment
Article Snippet: .. Immunofluorescence Mature CD4SP thymocytes were sorted and allowed to adhere to poly-L-lysine-coated glass slides (P0425; Sigma–Aldrich, St. Louis, MO). .. Cells were then fixed with 4% PFA in PBS and permeabilized with 0.1% Triton X-100 for 10 min on ice, followed by blocking (5% normal goat serum, 2% BSA in PBS) and staining (2% goat serum, 0.1% BSA, 0.1% Tween-20 in PBS).

Article Title: Necessity and Contingency in Developmental Genetic Screens: EGF, Wnt, and Semaphorin Pathways in Vulval Induction of the Nematode Oscheius tipulae
Article Snippet: Paragraph title: Immunofluorescence staining ... In brief, OP50 -grown populations were washed three times in distilled water and placed onto poly-l -lysine-coated (P0425-72EA; Sigma [Sigma Chemical], St. Louis, MO) slides prior to freeze-cracking.

Immunohistochemistry:

Article Title: Modulation of stress proteins and apoptotic regulators in the anoxia tolerant turtle brain
Article Snippet: .. The immunohistochemical analysis included regions of the turtle brain between levels A4.8 and A3.2 as defined by , e.g. the cortex (cortex medialis, dorsalis and pyriforme), pallial thickening, primordium hippocampi, dorsal ventricular ridge (DVR) of the telencephalon including the core nucleus, paleostriatum augmentatum, globus pallidus, and ventral paleostriatum Other sections were floated onto poly-L-lysine coated slides (Poly-PrepTM slides, Sigma P0425-72EA) and air-dried for cresyl violet staining. .. Immunohistochemical labeling was performed using a free-floating method in which relatively thick sections were reproducibly exposed to fixed volumes of antibody solutions ( ; ).

Microscopy:

Article Title: Study of amyloids using yeast
Article Snippet: .. To further reduce movement microscope slides can be coated with poly l-lysine (Sigma P0425) or the lectin concanavalin A, which both bind the yeast cell wall. .. Cover slips cleaned by soaking in 1 M NaOH and rinsed with distilled water can be coated with a 2 mg/ml solution of concanavalin A (Sigma C-5275) for 10 minutes.

Purification:

Article Title: Myeloid progenitor cluster formation drives emergency and leukemic myelopoiesis
Article Snippet: .. Immunofluorescence on purified cells GMP (2,000 cells/slide) were directly sorted onto poly-lysine coated slides (Sigma-Aldrich, P0425-72EA) and let settle down for 15 min at RT. .. Cells were washed 3x for 5 min with PBS at RT between each staining step.

Metabolic Labelling:

Article Title: Dynamics of genome reorganization during human cardiogenesis reveal an RBM20-dependent splicing factory
Article Snippet: CM were metabolically selected by 4 days of culture in DMEM without glucose and pyruvate but supplemented with 4 μM lactate. .. Poly-Lysine slides (Sigma-Aldrich, P0425) were pre-coated with 0.5 or 2 μg mL−1 rhLaminin521 (for hESCs or CMs, respectively) dispersed on a ~2 cm2 spot outlined by a hydrophobic pen for 2 h at 37 °C.

Blocking Assay:

Article Title: MC-PPEA as a new and more potent inhibitor of CLP-induced sepsis and pulmonary inflammation than FK866
Article Snippet: Immunofluorescence imaging of nuclear translocation of NFκB in A549 cells A549 cells were seeded on poly-L-lysine-coated glass slides (Catalog#: p0425; Sigma-Aldrich) and cultured in a 35 mm Petri dish with 3 mL of media at 37°C in a humidified atmosphere of 5% CO2 . .. Afterward, the slides were rinsed three times in 2 mL of PBS for 5 min each, followed by 60 min in blocking buffer (PBS/5% normal goat serum; Catalog#: 5245; Cell Signaling Technology).

Article Title: Zinc finger protein Zfp335 is required for the formation of the naïve T cell compartment
Article Snippet: Immunofluorescence Mature CD4SP thymocytes were sorted and allowed to adhere to poly-L-lysine-coated glass slides (P0425; Sigma–Aldrich, St. Louis, MO). .. Cells were then fixed with 4% PFA in PBS and permeabilized with 0.1% Triton X-100 for 10 min on ice, followed by blocking (5% normal goat serum, 2% BSA in PBS) and staining (2% goat serum, 0.1% BSA, 0.1% Tween-20 in PBS).

In Situ Hybridization:

Article Title: The QTL GNP1 Encodes GA20ox1, Which Increases Grain Number and Yield by Increasing Cytokinin Activity in Rice Panicle Meristems
Article Snippet: Paragraph title: In Situ Hybridization ... The samples were then embedded in Paraplast Plus (Sigma, P3683), cut into 10 μm sections and mounted on pre-coated poly-prep slides (Sigma, P0425).

Plasmid Preparation:

Article Title: Novel initiator caspase reporters uncover previously unknown features of caspase-activating cells
Article Snippet: In all of the experiments, 1 µg of plasmid was transfected using Effectene (Qiagen) reagent, following manufacturer's instructions. .. In most experiments, cells were grown on normal coverslips placed in 6-well plates; however, coverslips coated with poly-lysine (Sigma, P0425-72EA) were used to facilitate the visualisation of cell projections and active actin in Fig. S1E .

Article Title: Novel initiator caspase reporters uncover previously unknown features of caspase-activating cells
Article Snippet: In all of the experiments, 1 µg of plasmid was transfected using Effectene (Qiagen) reagent, following manufacturer's instructions. .. In most experiments, cells were grown on normal coverslips placed in 6-well plates; however, coverslips coated with poly-lysine (Sigma, P0425-72EA) were used to facilitate the visualisation of cell projections and active actin in .

Selection:

Article Title: Dynamics of genome reorganization during human cardiogenesis reveal an RBM20-dependent splicing factory
Article Snippet: This well-established procedure efficiently removed contaminating non-CM cells (which do not tolerate lactate metabolic selection), thus highly enriching for CM ( > 95% as assessed by Flow Cytometry for cTnT and immunofluorescence for α-actinin). .. Poly-Lysine slides (Sigma-Aldrich, P0425) were pre-coated with 0.5 or 2 μg mL−1 rhLaminin521 (for hESCs or CMs, respectively) dispersed on a ~2 cm2 spot outlined by a hydrophobic pen for 2 h at 37 °C.

Sample Prep:

Article Title: Study of amyloids using yeast
Article Snippet: Paragraph title: 3.2 Microscopy and sample preparation ... To further reduce movement microscope slides can be coated with poly l-lysine (Sigma P0425) or the lectin concanavalin A, which both bind the yeast cell wall.

Concentration Assay:

Article Title: Study of amyloids using yeast
Article Snippet: In order to minimize movement of the yeast cells it is important to place them at relatively high concentration (clearly turbid) in a minimal volume (2 microliters) on a microscope slide. .. To further reduce movement microscope slides can be coated with poly l-lysine (Sigma P0425) or the lectin concanavalin A, which both bind the yeast cell wall.

Staining:

Article Title: Adrenergic nerve degeneration in bone marrow drives aging of the hematopoietic stem cell niche
Article Snippet: Paragraph title: Immunofluorescence staining and imaging of sorted HSCs ... Lineage− CD48− Sca-1+ c-Kit+ CD150+ HSCs were directly sorted on poly-lysine coated slides (P0425-72AE; Sigma), 500–2000 cells per slide, incubated for 10 min, fixed with 4% PFA for 10 min at room temperature, and permeabilized with 0.2% Triton X-100 for 20 min at room temperature.

Article Title: Engineering a haematopoietic stem cell niche by revitalizing mesenchymal stromal cells
Article Snippet: Paragraph title: Immunofluorescence staining and imaging of sorted HSCs ... After co-culture in serum-free medium supplemented with SCF (20ng/ml) and TPO (10ng/ml) for 6 days, Lin− Sca-1+ c-Kit+ CD48− CD150+ HSCs were directly sorted on polylysine-coated slides (P0425–72AE; Sigma), 500–2000 cells per slide, incubated for 10 min, fixed with 4 % PFA for 10 min at room temperature, and permeabilised with 0.2 % Triton X-100 for 20 min at room temperature.

Article Title: Modulation of stress proteins and apoptotic regulators in the anoxia tolerant turtle brain
Article Snippet: .. The immunohistochemical analysis included regions of the turtle brain between levels A4.8 and A3.2 as defined by , e.g. the cortex (cortex medialis, dorsalis and pyriforme), pallial thickening, primordium hippocampi, dorsal ventricular ridge (DVR) of the telencephalon including the core nucleus, paleostriatum augmentatum, globus pallidus, and ventral paleostriatum Other sections were floated onto poly-L-lysine coated slides (Poly-PrepTM slides, Sigma P0425-72EA) and air-dried for cresyl violet staining. .. Immunohistochemical labeling was performed using a free-floating method in which relatively thick sections were reproducibly exposed to fixed volumes of antibody solutions ( ; ).

Article Title: Myeloid progenitor cluster formation drives emergency and leukemic myelopoiesis
Article Snippet: Immunofluorescence on purified cells GMP (2,000 cells/slide) were directly sorted onto poly-lysine coated slides (Sigma-Aldrich, P0425-72EA) and let settle down for 15 min at RT. .. Cells were washed 3x for 5 min with PBS at RT between each staining step.

Article Title: Zinc finger protein Zfp335 is required for the formation of the naïve T cell compartment
Article Snippet: Immunofluorescence Mature CD4SP thymocytes were sorted and allowed to adhere to poly-L-lysine-coated glass slides (P0425; Sigma–Aldrich, St. Louis, MO). .. Cells were then fixed with 4% PFA in PBS and permeabilized with 0.1% Triton X-100 for 10 min on ice, followed by blocking (5% normal goat serum, 2% BSA in PBS) and staining (2% goat serum, 0.1% BSA, 0.1% Tween-20 in PBS).

Article Title: Necessity and Contingency in Developmental Genetic Screens: EGF, Wnt, and Semaphorin Pathways in Vulval Induction of the Nematode Oscheius tipulae
Article Snippet: Paragraph title: Immunofluorescence staining ... In brief, OP50 -grown populations were washed three times in distilled water and placed onto poly-l -lysine-coated (P0425-72EA; Sigma [Sigma Chemical], St. Louis, MO) slides prior to freeze-cracking.

Fluorescence In Situ Hybridization:

Article Title: Dynamics of genome reorganization during human cardiogenesis reveal an RBM20-dependent splicing factory
Article Snippet: Paragraph title: 3D DNA FISH ... Poly-Lysine slides (Sigma-Aldrich, P0425) were pre-coated with 0.5 or 2 μg mL−1 rhLaminin521 (for hESCs or CMs, respectively) dispersed on a ~2 cm2 spot outlined by a hydrophobic pen for 2 h at 37 °C.

other:

Article Title: Label-free quantitation of glycated hemoglobin in single red blood cells by transient absorption microscopy and phasor analysis
Article Snippet: Then, 1.5 μl of the whole blood was pipetted onto a poly-l -lysine–coated cover slide and sandwiched between a cover slide (0.17-mm thickness; VWR Micro Cover Glasses, 48393-048) and poly-l -lysine–coated cover slide (P0425-72 EA, Sigma-Aldrich).

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    Millipore poly d lysine
    Poly D Lysine, supplied by Millipore, used in various techniques. Bioz Stars score: 90/100, based on 183 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore anti acetylated lysine antibody
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