trypsin edta  (Thermo Fisher)


Bioz Verified Symbol Thermo Fisher is a verified supplier
Bioz Manufacturer Symbol Thermo Fisher manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    Name:
    Trypsin EDTA Solution TE
    Description:
    Gibco Trypsin solutions are made from trypsin powder a mixture of proteases derived from bovine pancreas Due to its digestive strength it is widely used for cell dissociation during routine cell culture passaging and primary tissue dissociation This Gibco Trypsin EDTA solution is formulation as 0 025 trypsin and 0 01 EDTA in Phosphate Buffered Saline PBS We offer a variety of Gibco Trypsin and animal origin free TrypLE cell dissociation products to meet your needs This trypsin is manufactured as follows WithEDTAPhenol RedThe complete formulation is available Product UseFor Research Use Only Not for use in diagnostic procedures cGMP Manufacturing and Quality SystemGibco Trypsin is manufactured at a cGMP compliant facility located in Grand Island New York The facility is registered with the FDA as a medical device manufacturer and is certified to ISO 13485 standards
    Catalog Number:
    R001100
    Price:
    None
    Category:
    Cell Culture Transfection Reagents
    Applications:
    Cell Culture|Corneal Epithelial Cell Culture|Fibroblast Culture|Keratinocyte Culture|Large Vessel Endothelial Cell Culture|Mammalian Cell Culture|Mammary Epithelial Cell Culture|Melanocyte Culture|Microvascular Endothelial Cell Culture|Smooth Muscle Cell Culture|Primary Cell Culture
    Buy from Supplier


    Structured Review

    Thermo Fisher trypsin edta
    Gibco Trypsin solutions are made from trypsin powder a mixture of proteases derived from bovine pancreas Due to its digestive strength it is widely used for cell dissociation during routine cell culture passaging and primary tissue dissociation This Gibco Trypsin EDTA solution is formulation as 0 025 trypsin and 0 01 EDTA in Phosphate Buffered Saline PBS We offer a variety of Gibco Trypsin and animal origin free TrypLE cell dissociation products to meet your needs This trypsin is manufactured as follows WithEDTAPhenol RedThe complete formulation is available Product UseFor Research Use Only Not for use in diagnostic procedures cGMP Manufacturing and Quality SystemGibco Trypsin is manufactured at a cGMP compliant facility located in Grand Island New York The facility is registered with the FDA as a medical device manufacturer and is certified to ISO 13485 standards
    https://www.bioz.com/result/trypsin edta/product/Thermo Fisher
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    trypsin edta - by Bioz Stars, 2021-04
    99/100 stars

    Images

    Related Articles

    Infection:

    Article Title: Human Rotavirus VP6-Specific Antibodies Mediate Intracellular Neutralization by Binding to a Quaternary Structure in the Transcriptional Pore
    Article Snippet: Cells MA-104 Clone 1 cells (ATCC CRL-2378.1) and Madin Darbin canine kidney (MDCK) cells (ATCC CCL-34) were grown in complete medium (CM) consisting of DMEM with 4.5% glucose (Mediatech) supplemented with 10% (v/v) fetal bovine serum (Invitrogen), 0.1 mM MEM non-essential amino acids (Invitrogen), 100 I.U./mL penicillin, 100 µg/mL streptomycin, 2 mM glutamine and 1 mM sodium pyruvate (all from Mediatech). .. Infection medium (IM), used when cells were inoculated with rotavirus and cultured, contained all of the above supplements except serum, and trypsin-EDTA (Invitrogen) was added to a final concentration of 1 µg/mL. .. A lineage of Caco-2 cells (a kind gift from Dr. Blaise Corthésy) derived by passage from a commercial cell line (HTB-37, American Type Culture Collection) were grown in CM supplemented with 0.1% transferrin (Sigma).

    Cell Culture:

    Article Title: Human Rotavirus VP6-Specific Antibodies Mediate Intracellular Neutralization by Binding to a Quaternary Structure in the Transcriptional Pore
    Article Snippet: Cells MA-104 Clone 1 cells (ATCC CRL-2378.1) and Madin Darbin canine kidney (MDCK) cells (ATCC CCL-34) were grown in complete medium (CM) consisting of DMEM with 4.5% glucose (Mediatech) supplemented with 10% (v/v) fetal bovine serum (Invitrogen), 0.1 mM MEM non-essential amino acids (Invitrogen), 100 I.U./mL penicillin, 100 µg/mL streptomycin, 2 mM glutamine and 1 mM sodium pyruvate (all from Mediatech). .. Infection medium (IM), used when cells were inoculated with rotavirus and cultured, contained all of the above supplements except serum, and trypsin-EDTA (Invitrogen) was added to a final concentration of 1 µg/mL. .. A lineage of Caco-2 cells (a kind gift from Dr. Blaise Corthésy) derived by passage from a commercial cell line (HTB-37, American Type Culture Collection) were grown in CM supplemented with 0.1% transferrin (Sigma).

    Concentration Assay:

    Article Title: Human Rotavirus VP6-Specific Antibodies Mediate Intracellular Neutralization by Binding to a Quaternary Structure in the Transcriptional Pore
    Article Snippet: Cells MA-104 Clone 1 cells (ATCC CRL-2378.1) and Madin Darbin canine kidney (MDCK) cells (ATCC CCL-34) were grown in complete medium (CM) consisting of DMEM with 4.5% glucose (Mediatech) supplemented with 10% (v/v) fetal bovine serum (Invitrogen), 0.1 mM MEM non-essential amino acids (Invitrogen), 100 I.U./mL penicillin, 100 µg/mL streptomycin, 2 mM glutamine and 1 mM sodium pyruvate (all from Mediatech). .. Infection medium (IM), used when cells were inoculated with rotavirus and cultured, contained all of the above supplements except serum, and trypsin-EDTA (Invitrogen) was added to a final concentration of 1 µg/mL. .. A lineage of Caco-2 cells (a kind gift from Dr. Blaise Corthésy) derived by passage from a commercial cell line (HTB-37, American Type Culture Collection) were grown in CM supplemented with 0.1% transferrin (Sigma).

    Modification:

    Article Title: Grb7-derived calmodulin-binding peptides inhibit proliferation, migration and invasiveness of tumor cells while they enhance attachment to the substrate
    Article Snippet: .. Dulbecco's modified Eagle's medium (DMEM), trypsin/EDTA, L-glutamine and FBS were obtained from Gibco; Anti-mouse IgG (whole molecule) HRP-conjugated secondary antibody (A9044), phenyl-Sepharose 6 (fast-flow), dansyl chloride, glutaraldehyde, N,N,N′,N′ -tetramethylethylenediamine, Triton X-100, Fast Green FC and Crystal Violet were purchased from Sigma-Aldrich; polyvinylidene difluoride (PVDF) membranes, the mouse monoclonal anti-phosphotyrosine antibody (clone 4G10, IgG2bκ isotype) made against phosphotyramine-KLH (keyhole limpet hemocyanin), fibronectin, dimethyl sulfoxide (DMSO), acrylamide and EGF were from Merck Millipore; W-7 and W-12 were obtained from Calbiochem or Abcam, and 5(6)-TAMRA was from Novabiochem®. .. The Diff-Quik staining kit was from Medion Diagnostics AG, bovine serum albumin (BSA) was from Nzytech and the enhanced chemiluminescence (ECL) kit was purchased from GE Healthcare-Amersham.

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 97
    Thermo Fisher u251
    Projection electrophoresis permits the simultaneous analysis of hundreds of single cells by concurrent separation after simultaneous lysis. ( a ) Maximum intensity projection 3D renderings of example separation lanes read out by tiled light-sheet microscopy. ( b ) xz ( y -summed) contour plots of background-subtracted actinin and GAPDH protein signal within the lanes depicted in ( a ). ( c ) Revolved z -intensity profiles (arbitrary fluorescence units, AFU, vs. z ) for the four separation lanes depicted in ( a , b ). Each plot depicts z -directional intensity profiles (summed fluorescence within each xy ROI vs. z ) for GAPDH (magenta) and actinin (cyan), revolved around the z -axis to generate 3D rendering of fluorescence distribution. ( d ) Histogram quantification shows 86% of <t>U251</t> cells lyse within 5 s of initiating lysis. ( e ) Quantified fluorescence intensity data for n = 159 separation lanes passing quality control for both the GAPDH and actinin channels. Each plot depicts revolved z -intensity profiles. ( f ) Full-gel wide-field fluorescence image of calcein-stained live BT474 breast tumor cells before analysis. ( g ) Subset of the live cells from ( f ), within a 1.75 × 1.75 mm light-sheet microscopy field of view (scale bar depicts 200 μm). ( h ) Post-separation wide-field fluorescence image of probed GAPDH signal within the same field of view as ( g ). ( f – h ) are representative of n > 3 separation gels. ( i ) Maximum intensity projection 3D rendering (representative of duplicate separation gels) of a light-sheet microscopy image (same field of view as ( g , h )), showing 3D separations of GAPDH and actinin from tens of separation lanes, each corresponding to signal from the settled cells in microwells depicted in ( f , g ). ( j ) Overlay image of segmented spots corresponding to live BT474 cells in microwells (green) and probed GAPDH bands after separation (magenta), for the same separation gel depicted in ( f , i ). ( k ) Quantification of correspondence between the segmented live cells and bands (via intensity thresholding) within the same separation lanes as ( j ). ( l ) Quantified migration distances from a total of n = 507 (GAPDH) and n = 303 (actinin) lanes passing quality control in two projection electrophoresis gels. ( m ) Quantified z -direction peak widths for the same bands analyzed in ( l ). (n-o) Map of the variation in GAPDH ( n ) and actinin ( o ) electromigration distances across the xy gel area. Source data are provided as a Source Data file.
    U251, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/u251/product/Thermo Fisher
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    u251 - by Bioz Stars, 2021-04
    97/100 stars
      Buy from Supplier

    97
    Thermo Fisher pc14 pe6 pgf1 br4 cells
    Nintedanib and anti-VEGF/Ang2 nanobody prevent brain metastases formation. 9.4 T MRI after Gadolinium contrast administration was performed on day 26 after intracardial tumor cell injection. a Bar chart illustrating the reduced percentage of mice with metastases, detectable in cMRI. b Representative T1-w cMRI images depicting larger intracranial metastases in the control groups, indicated by arrowheads. scale bar = 2 mm. c Scatter plots showing the reduction of number and volume of cranial metastases in cMRI. d Scatter plots depicting the number and volume of meningeal metastases in cMRI e Quantification demonstrating a higher histological tumor–tissue ratio in control mice at their time of death. f Representative histological slices with higher number and size of <t>PC14-PE6</t> <t>pGF1</t> <t>Br4</t> metastases in the control group. Fluorescent staining: DAPI (blue) = nucleus, GFP (green) = PC14-PE6 tumor cells, Alexa Flour 546 (orange) = collagen-IV positive vascular basement membrane. Arrows indicate GFP-positive metastatic lesions. Scale bar = 1000 µm. Mean values with standard errors of the mean are shown. *p
    Pc14 Pe6 Pgf1 Br4 Cells, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pc14 pe6 pgf1 br4 cells/product/Thermo Fisher
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    pc14 pe6 pgf1 br4 cells - by Bioz Stars, 2021-04
    97/100 stars
      Buy from Supplier

    97
    Thermo Fisher trypsin edta
    Membrane binding of the key PNN component aggrecan is biochemically altered in <t>Ptprz1</t> KO and Tnr KO mice brains. Brain homogenates were treated with ChABC to remove the hyaluronan backbone of PNNs or chondroitinase in the presence of <t>EDTA</t> (ChABC EDTA) and centrifuged to obtain soluble release ( R ) and insoluble pellet ( P ) fractions. A , Western blotting image showing release of PNN marker aggrecan into soluble phase from brain homogenates of WT, Ptprz1 KO, and Tnr KO mice by ChABC treatment alone and ChABC EDTA treatment. The release of aggrecan into the soluble fraction required ChABC treatment in addition with EDTA in WT brain homogenates. Aggrecan was released more readily with just ChABC treatment in Ptprz1 KO and Tnr KO animals. B , quantification showing ratio of the soluble release fraction ( R ) to total aggrecan levels (soluble release (R) + insoluble pellet (P)) in WT, Ptprz1 KO, and Tnr KO mice. There was a statistically significant difference in the release of aggrecan among the three genotypes as determined by one-way ANOVA (F(2,7) = 14.94, p = 0.0030). A Tukey's post hoc test showed that aggrecan was released much more readily in Ptprz1 KO mice (42 ± 0.04%, p = 0.0340) as well as in Tnr KO mice (58 ± 13%, p = 0.0024) compared with WT mice (15 ± 9%) when treated with just ChABC. There was no significant difference in aggrecan release between Ptprz1 KO and Tnr KO brains. Treatment with ChABC alongside EDTA led to almost complete release of aggrecan into the soluble release fraction in all genotypes. The ratio of release to total in Ptprz1 KO, Tnr KO and WT mice was not significantly different among the genotypes in the ChABC with EDTA treatment group. B , bars in graphs represent percentage release ± S.D.
    Trypsin Edta, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/trypsin edta/product/Thermo Fisher
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    trypsin edta - by Bioz Stars, 2021-04
    97/100 stars
      Buy from Supplier

    Image Search Results


    Projection electrophoresis permits the simultaneous analysis of hundreds of single cells by concurrent separation after simultaneous lysis. ( a ) Maximum intensity projection 3D renderings of example separation lanes read out by tiled light-sheet microscopy. ( b ) xz ( y -summed) contour plots of background-subtracted actinin and GAPDH protein signal within the lanes depicted in ( a ). ( c ) Revolved z -intensity profiles (arbitrary fluorescence units, AFU, vs. z ) for the four separation lanes depicted in ( a , b ). Each plot depicts z -directional intensity profiles (summed fluorescence within each xy ROI vs. z ) for GAPDH (magenta) and actinin (cyan), revolved around the z -axis to generate 3D rendering of fluorescence distribution. ( d ) Histogram quantification shows 86% of U251 cells lyse within 5 s of initiating lysis. ( e ) Quantified fluorescence intensity data for n = 159 separation lanes passing quality control for both the GAPDH and actinin channels. Each plot depicts revolved z -intensity profiles. ( f ) Full-gel wide-field fluorescence image of calcein-stained live BT474 breast tumor cells before analysis. ( g ) Subset of the live cells from ( f ), within a 1.75 × 1.75 mm light-sheet microscopy field of view (scale bar depicts 200 μm). ( h ) Post-separation wide-field fluorescence image of probed GAPDH signal within the same field of view as ( g ). ( f – h ) are representative of n > 3 separation gels. ( i ) Maximum intensity projection 3D rendering (representative of duplicate separation gels) of a light-sheet microscopy image (same field of view as ( g , h )), showing 3D separations of GAPDH and actinin from tens of separation lanes, each corresponding to signal from the settled cells in microwells depicted in ( f , g ). ( j ) Overlay image of segmented spots corresponding to live BT474 cells in microwells (green) and probed GAPDH bands after separation (magenta), for the same separation gel depicted in ( f , i ). ( k ) Quantification of correspondence between the segmented live cells and bands (via intensity thresholding) within the same separation lanes as ( j ). ( l ) Quantified migration distances from a total of n = 507 (GAPDH) and n = 303 (actinin) lanes passing quality control in two projection electrophoresis gels. ( m ) Quantified z -direction peak widths for the same bands analyzed in ( l ). (n-o) Map of the variation in GAPDH ( n ) and actinin ( o ) electromigration distances across the xy gel area. Source data are provided as a Source Data file.

    Journal: Nature Communications

    Article Title: 3D projection electrophoresis for single-cell immunoblotting

    doi: 10.1038/s41467-020-19738-1

    Figure Lengend Snippet: Projection electrophoresis permits the simultaneous analysis of hundreds of single cells by concurrent separation after simultaneous lysis. ( a ) Maximum intensity projection 3D renderings of example separation lanes read out by tiled light-sheet microscopy. ( b ) xz ( y -summed) contour plots of background-subtracted actinin and GAPDH protein signal within the lanes depicted in ( a ). ( c ) Revolved z -intensity profiles (arbitrary fluorescence units, AFU, vs. z ) for the four separation lanes depicted in ( a , b ). Each plot depicts z -directional intensity profiles (summed fluorescence within each xy ROI vs. z ) for GAPDH (magenta) and actinin (cyan), revolved around the z -axis to generate 3D rendering of fluorescence distribution. ( d ) Histogram quantification shows 86% of U251 cells lyse within 5 s of initiating lysis. ( e ) Quantified fluorescence intensity data for n = 159 separation lanes passing quality control for both the GAPDH and actinin channels. Each plot depicts revolved z -intensity profiles. ( f ) Full-gel wide-field fluorescence image of calcein-stained live BT474 breast tumor cells before analysis. ( g ) Subset of the live cells from ( f ), within a 1.75 × 1.75 mm light-sheet microscopy field of view (scale bar depicts 200 μm). ( h ) Post-separation wide-field fluorescence image of probed GAPDH signal within the same field of view as ( g ). ( f – h ) are representative of n > 3 separation gels. ( i ) Maximum intensity projection 3D rendering (representative of duplicate separation gels) of a light-sheet microscopy image (same field of view as ( g , h )), showing 3D separations of GAPDH and actinin from tens of separation lanes, each corresponding to signal from the settled cells in microwells depicted in ( f , g ). ( j ) Overlay image of segmented spots corresponding to live BT474 cells in microwells (green) and probed GAPDH bands after separation (magenta), for the same separation gel depicted in ( f , i ). ( k ) Quantification of correspondence between the segmented live cells and bands (via intensity thresholding) within the same separation lanes as ( j ). ( l ) Quantified migration distances from a total of n = 507 (GAPDH) and n = 303 (actinin) lanes passing quality control in two projection electrophoresis gels. ( m ) Quantified z -direction peak widths for the same bands analyzed in ( l ). (n-o) Map of the variation in GAPDH ( n ) and actinin ( o ) electromigration distances across the xy gel area. Source data are provided as a Source Data file.

    Article Snippet: Single-cell separations Adherent U251 glioblastoma and BT474 breast tumor cells were detached from culture flasks with 0.05% Trypsin-EDTA (Life Technologies 25300120) (U251) or 5 mM EDTA (Invitrogen 15575-038) in PBS (BT474) and resuspended in cold PBS (Life Technologies 10010049) at a concentration of 1.5 × 106 cells/mL.

    Techniques: Electrophoresis, Lysis, Microscopy, Fluorescence, Staining, Migration

    Design and verification of sample preparation for projection electrophoresis of single mammalian cells. ( a ) High-density endogenous protein bands (ii) correspond to single-cell settling in microwells (i). Scale bars represent 1 mm (left full-gel images) and 200 μm (right zoom images). ( b ) Illustration of top-view and side-view geometries shown in protein dilution studies ( c , d ). ( c ) Modeling and experimental quantification of diffusional dilution during lysis. Simulated and experimental top-view images of diffusional protein dilution during lysis, and side-view simulated results are shown. The simulated initial TurboGFP concentration was 2 μM. Experimental image is representative of 12 monitored cells across 3 independent lysis experiments. Scale bars represent 50 μm. ( d ) Modeling the impact of diffusion during electrophoresis on detectable in-gel protein concentration. Side and top view TurboGFP concentration profiles are shown at different times during electrophoresis. Simulated initial TurboGFP concentration (before lysis and electrophoresis) was 2 μM. Scale bars represent 30 μm. ( e ) Quantification of the percent of protein remaining in the microwell region during lysis (experiment plots mean and standard deviation of n = 12 cells across 3 lysis experiments). ( f ) Quantification of the change in the spatial maximum protein concentration as a function of time after protein solubilization (experiment plots mean and standard deviation of n = 12 cells across 3 lysis experiments). ( g ) Simulated maximum protein concentration vs. electrophoresis time, for 3 model proteins. ( h ) Representative β-tubulin separations from U251 glioblastoma cells lysed with different buffer formulations (2× RIPA lysis buffer and 2× RIPA including 8 M urea), both after 10 s electrophoresis. Lysis/EP buffer requires 8 M urea for fast protein solubilization and electromigration. ( i ) Maximum intensity projection 3D renderings and z -intensity profiles of probed GAPDH bands from single BT474 breast tumor cells. ( j ) Microwell packing density (impacting assay throughput) is dependent on protein band diffusion before photocapture. Protein diffusion profiles confirm that a microwell pitch of 200 μm is sufficient to resolve bands from neighboring microwells. After 10 s EP, the mean peak width ( σ xy ) of the xy GAPDH spots is 32 ± 11 μm (mean ± standard deviation of n = 47 single-cell separation lanes across 5 replicate separation gels). The depicted confocal slice micrograph is representative of 20 confocal image stacks, across different regions of 5 replicate separation gels. At a microwell pitch of 192 μm (6 σ xy ),

    Journal: Nature Communications

    Article Title: 3D projection electrophoresis for single-cell immunoblotting

    doi: 10.1038/s41467-020-19738-1

    Figure Lengend Snippet: Design and verification of sample preparation for projection electrophoresis of single mammalian cells. ( a ) High-density endogenous protein bands (ii) correspond to single-cell settling in microwells (i). Scale bars represent 1 mm (left full-gel images) and 200 μm (right zoom images). ( b ) Illustration of top-view and side-view geometries shown in protein dilution studies ( c , d ). ( c ) Modeling and experimental quantification of diffusional dilution during lysis. Simulated and experimental top-view images of diffusional protein dilution during lysis, and side-view simulated results are shown. The simulated initial TurboGFP concentration was 2 μM. Experimental image is representative of 12 monitored cells across 3 independent lysis experiments. Scale bars represent 50 μm. ( d ) Modeling the impact of diffusion during electrophoresis on detectable in-gel protein concentration. Side and top view TurboGFP concentration profiles are shown at different times during electrophoresis. Simulated initial TurboGFP concentration (before lysis and electrophoresis) was 2 μM. Scale bars represent 30 μm. ( e ) Quantification of the percent of protein remaining in the microwell region during lysis (experiment plots mean and standard deviation of n = 12 cells across 3 lysis experiments). ( f ) Quantification of the change in the spatial maximum protein concentration as a function of time after protein solubilization (experiment plots mean and standard deviation of n = 12 cells across 3 lysis experiments). ( g ) Simulated maximum protein concentration vs. electrophoresis time, for 3 model proteins. ( h ) Representative β-tubulin separations from U251 glioblastoma cells lysed with different buffer formulations (2× RIPA lysis buffer and 2× RIPA including 8 M urea), both after 10 s electrophoresis. Lysis/EP buffer requires 8 M urea for fast protein solubilization and electromigration. ( i ) Maximum intensity projection 3D renderings and z -intensity profiles of probed GAPDH bands from single BT474 breast tumor cells. ( j ) Microwell packing density (impacting assay throughput) is dependent on protein band diffusion before photocapture. Protein diffusion profiles confirm that a microwell pitch of 200 μm is sufficient to resolve bands from neighboring microwells. After 10 s EP, the mean peak width ( σ xy ) of the xy GAPDH spots is 32 ± 11 μm (mean ± standard deviation of n = 47 single-cell separation lanes across 5 replicate separation gels). The depicted confocal slice micrograph is representative of 20 confocal image stacks, across different regions of 5 replicate separation gels. At a microwell pitch of 192 μm (6 σ xy ),

    Article Snippet: Single-cell separations Adherent U251 glioblastoma and BT474 breast tumor cells were detached from culture flasks with 0.05% Trypsin-EDTA (Life Technologies 25300120) (U251) or 5 mM EDTA (Invitrogen 15575-038) in PBS (BT474) and resuspended in cold PBS (Life Technologies 10010049) at a concentration of 1.5 × 106 cells/mL.

    Techniques: Sample Prep, Electrophoresis, Lysis, Concentration Assay, Diffusion-based Assay, Protein Concentration, Standard Deviation

    Nintedanib and anti-VEGF/Ang2 nanobody prevent brain metastases formation. 9.4 T MRI after Gadolinium contrast administration was performed on day 26 after intracardial tumor cell injection. a Bar chart illustrating the reduced percentage of mice with metastases, detectable in cMRI. b Representative T1-w cMRI images depicting larger intracranial metastases in the control groups, indicated by arrowheads. scale bar = 2 mm. c Scatter plots showing the reduction of number and volume of cranial metastases in cMRI. d Scatter plots depicting the number and volume of meningeal metastases in cMRI e Quantification demonstrating a higher histological tumor–tissue ratio in control mice at their time of death. f Representative histological slices with higher number and size of PC14-PE6 pGF1 Br4 metastases in the control group. Fluorescent staining: DAPI (blue) = nucleus, GFP (green) = PC14-PE6 tumor cells, Alexa Flour 546 (orange) = collagen-IV positive vascular basement membrane. Arrows indicate GFP-positive metastatic lesions. Scale bar = 1000 µm. Mean values with standard errors of the mean are shown. *p

    Journal: Clinical & Experimental Metastasis

    Article Title: Nintedanib and a bi-specific anti-VEGF/Ang2 nanobody selectively prevent brain metastases of lung adenocarcinoma cells

    doi: 10.1007/s10585-020-10055-x

    Figure Lengend Snippet: Nintedanib and anti-VEGF/Ang2 nanobody prevent brain metastases formation. 9.4 T MRI after Gadolinium contrast administration was performed on day 26 after intracardial tumor cell injection. a Bar chart illustrating the reduced percentage of mice with metastases, detectable in cMRI. b Representative T1-w cMRI images depicting larger intracranial metastases in the control groups, indicated by arrowheads. scale bar = 2 mm. c Scatter plots showing the reduction of number and volume of cranial metastases in cMRI. d Scatter plots depicting the number and volume of meningeal metastases in cMRI e Quantification demonstrating a higher histological tumor–tissue ratio in control mice at their time of death. f Representative histological slices with higher number and size of PC14-PE6 pGF1 Br4 metastases in the control group. Fluorescent staining: DAPI (blue) = nucleus, GFP (green) = PC14-PE6 tumor cells, Alexa Flour 546 (orange) = collagen-IV positive vascular basement membrane. Arrows indicate GFP-positive metastatic lesions. Scale bar = 1000 µm. Mean values with standard errors of the mean are shown. *p

    Article Snippet: Briefly, PC14-PE6 pGF1 Br4 cells were trypsinized (Gibco, Life Science Technologies, cat. no.: 25200-056), washed, counted and resuspended in PBS (cat. no: D8537, Sigma Life Sciences) in a final concentration of 5 × 106 cells/mL.

    Techniques: Magnetic Resonance Imaging, Injection, Mouse Assay, Staining

    Membrane binding of the key PNN component aggrecan is biochemically altered in Ptprz1 KO and Tnr KO mice brains. Brain homogenates were treated with ChABC to remove the hyaluronan backbone of PNNs or chondroitinase in the presence of EDTA (ChABC EDTA) and centrifuged to obtain soluble release ( R ) and insoluble pellet ( P ) fractions. A , Western blotting image showing release of PNN marker aggrecan into soluble phase from brain homogenates of WT, Ptprz1 KO, and Tnr KO mice by ChABC treatment alone and ChABC EDTA treatment. The release of aggrecan into the soluble fraction required ChABC treatment in addition with EDTA in WT brain homogenates. Aggrecan was released more readily with just ChABC treatment in Ptprz1 KO and Tnr KO animals. B , quantification showing ratio of the soluble release fraction ( R ) to total aggrecan levels (soluble release (R) + insoluble pellet (P)) in WT, Ptprz1 KO, and Tnr KO mice. There was a statistically significant difference in the release of aggrecan among the three genotypes as determined by one-way ANOVA (F(2,7) = 14.94, p = 0.0030). A Tukey's post hoc test showed that aggrecan was released much more readily in Ptprz1 KO mice (42 ± 0.04%, p = 0.0340) as well as in Tnr KO mice (58 ± 13%, p = 0.0024) compared with WT mice (15 ± 9%) when treated with just ChABC. There was no significant difference in aggrecan release between Ptprz1 KO and Tnr KO brains. Treatment with ChABC alongside EDTA led to almost complete release of aggrecan into the soluble release fraction in all genotypes. The ratio of release to total in Ptprz1 KO, Tnr KO and WT mice was not significantly different among the genotypes in the ChABC with EDTA treatment group. B , bars in graphs represent percentage release ± S.D.

    Journal: The Journal of Biological Chemistry

    Article Title: The protein tyrosine phosphatase RPTPζ/phosphacan is critical for perineuronal net structure

    doi: 10.1074/jbc.RA119.010830

    Figure Lengend Snippet: Membrane binding of the key PNN component aggrecan is biochemically altered in Ptprz1 KO and Tnr KO mice brains. Brain homogenates were treated with ChABC to remove the hyaluronan backbone of PNNs or chondroitinase in the presence of EDTA (ChABC EDTA) and centrifuged to obtain soluble release ( R ) and insoluble pellet ( P ) fractions. A , Western blotting image showing release of PNN marker aggrecan into soluble phase from brain homogenates of WT, Ptprz1 KO, and Tnr KO mice by ChABC treatment alone and ChABC EDTA treatment. The release of aggrecan into the soluble fraction required ChABC treatment in addition with EDTA in WT brain homogenates. Aggrecan was released more readily with just ChABC treatment in Ptprz1 KO and Tnr KO animals. B , quantification showing ratio of the soluble release fraction ( R ) to total aggrecan levels (soluble release (R) + insoluble pellet (P)) in WT, Ptprz1 KO, and Tnr KO mice. There was a statistically significant difference in the release of aggrecan among the three genotypes as determined by one-way ANOVA (F(2,7) = 14.94, p = 0.0030). A Tukey's post hoc test showed that aggrecan was released much more readily in Ptprz1 KO mice (42 ± 0.04%, p = 0.0340) as well as in Tnr KO mice (58 ± 13%, p = 0.0024) compared with WT mice (15 ± 9%) when treated with just ChABC. There was no significant difference in aggrecan release between Ptprz1 KO and Tnr KO brains. Treatment with ChABC alongside EDTA led to almost complete release of aggrecan into the soluble release fraction in all genotypes. The ratio of release to total in Ptprz1 KO, Tnr KO and WT mice was not significantly different among the genotypes in the ChABC with EDTA treatment group. B , bars in graphs represent percentage release ± S.D.

    Article Snippet: Briefly, cortices of embryonic day (E) 16 CD-1 WT or Ptprz1 KO embryos were removed and digested in 0.25% trypsin-EDTA (Thermo Fisher Scientific).

    Techniques: Binding Assay, Mouse Assay, Western Blot, Marker

    Signalling pathways in TL2i cells. ( a ) Phase-contrast microphotographs, AP activity and immunofluorescence labelling of OCT4, SSEA4 and TRA-1–60 in TL2i cells. ( b ) A colony-forming assay with TL2i-OS3–10 and TL2i-H9S3-2 cells (representative experiment). Cell clumps were plated in TL2i medium supplemented with pharmacological inhibitors of JAK2 (SD1029 at 10 μM), FGFR (SU5402 at 10 μM) and SMADs (SB431542 at 10 μM), and cultivated for 5 days. Upper panels: staining to reveal AP activity; bottom panels: histograms showing the percentage of undifferentiated, mixed and differentiated colonies. ( n =3; error bars indicates the mean±s.e.m.). ( c ) Histogram representation of the mRNA level (ΔCt) of pluripotency genes in TL2i-OS3–10 cells before and treatment with FGFR inhibitor SU5402 for 5 days after normalization to GAPDH (ΔCt=1). ( n =3, mean±s.d.). ( d ) Characteristics of TL2i-OS3–10 cells after propagation on Matrigel without MEF; AP, alkaline phosphatase activity. ( e ) Histograms showing the percentage of undifferentiated, mixed and differentiated colonies ( n =3; error bars indicate the mean±s.e.m.) in a colony-forming assay with TL2i-OS3–10 cells (representative experiment). Cell clumps were plated in a medium supplemented with pharmacological inhibitors of FGFR (SU5402 at 10 μM) and SMADs (SB431542 at 10 μM), and cultivated for 5 days with LIF+4′-OHT. ( f ) Histogram representation of the mRNA level (ΔCt) of LIFR , GP130 , JAK and STAT3 genes in TL2i-H9S3-2 cells, after normalization to GAPDH (ΔCt=1), then to TL-H9S3-2 (blue bars) and F-H9S3-2 cells (red bars). ( n =3, mean±s. d.). ( g ) Western blot analysis of STAT3 and STAT3-ER T2 expression in OSCAR, F-H9S3-2, TL-H9S3-2 and TL2i-H9S3-2 cells, analysed with antibodies to total STAT3, phospho-(Tyr705)-STAT3 and phospho-(Ser720)-STAT3. One representative experiment of three repeats is shown. ( h ) Phase-contrast microphotographs (PC) and immunofluorescence labelling of OCT4, NANOG, SSEA4 and TRA-1–81 in TL2i-OS3–10 cells after culturing in 2i/LIF medium without 4′-OHT for 30 passages. ( i ) Western blot analysis of STAT3 and STAT3-ER T2 expression in TL-OS3–10 cells and TL2i-OS3–10 cells (+/−4′-OHT), analysed with antibodies to total STAT3. One representative experiment of two repeats is shown. ( j ) Phase-contrast microphotographs (PC), AP detection and immunofluorescence labelling of OCT4, NANOG, TRA-1–81 and SSEA4 in TL2i-OS3–10 cells after culturing in N2B27+2i/LIF basal medium for eight passages. ( k ) Histogram representation of the cloning efficiency of F, TL and TL2i cells (OS3–10, H9S3–2 and H9S3–14 lines) after single-cell dissociation with 0.05% trypsin-EDTA and re-plating on feeders in the presence of 10 μM ROCK inhibitor Y-27632 for 24 h post dissociation. Tukey's test; n =3; error bars indicate the mean±s. e. Scale bar, 50 μm ( a , d , h , i ).

    Journal: Nature Communications

    Article Title: Reinforcement of STAT3 activity reprogrammes human embryonic stem cells to naive-like pluripotency

    doi: 10.1038/ncomms8095

    Figure Lengend Snippet: Signalling pathways in TL2i cells. ( a ) Phase-contrast microphotographs, AP activity and immunofluorescence labelling of OCT4, SSEA4 and TRA-1–60 in TL2i cells. ( b ) A colony-forming assay with TL2i-OS3–10 and TL2i-H9S3-2 cells (representative experiment). Cell clumps were plated in TL2i medium supplemented with pharmacological inhibitors of JAK2 (SD1029 at 10 μM), FGFR (SU5402 at 10 μM) and SMADs (SB431542 at 10 μM), and cultivated for 5 days. Upper panels: staining to reveal AP activity; bottom panels: histograms showing the percentage of undifferentiated, mixed and differentiated colonies. ( n =3; error bars indicates the mean±s.e.m.). ( c ) Histogram representation of the mRNA level (ΔCt) of pluripotency genes in TL2i-OS3–10 cells before and treatment with FGFR inhibitor SU5402 for 5 days after normalization to GAPDH (ΔCt=1). ( n =3, mean±s.d.). ( d ) Characteristics of TL2i-OS3–10 cells after propagation on Matrigel without MEF; AP, alkaline phosphatase activity. ( e ) Histograms showing the percentage of undifferentiated, mixed and differentiated colonies ( n =3; error bars indicate the mean±s.e.m.) in a colony-forming assay with TL2i-OS3–10 cells (representative experiment). Cell clumps were plated in a medium supplemented with pharmacological inhibitors of FGFR (SU5402 at 10 μM) and SMADs (SB431542 at 10 μM), and cultivated for 5 days with LIF+4′-OHT. ( f ) Histogram representation of the mRNA level (ΔCt) of LIFR , GP130 , JAK and STAT3 genes in TL2i-H9S3-2 cells, after normalization to GAPDH (ΔCt=1), then to TL-H9S3-2 (blue bars) and F-H9S3-2 cells (red bars). ( n =3, mean±s. d.). ( g ) Western blot analysis of STAT3 and STAT3-ER T2 expression in OSCAR, F-H9S3-2, TL-H9S3-2 and TL2i-H9S3-2 cells, analysed with antibodies to total STAT3, phospho-(Tyr705)-STAT3 and phospho-(Ser720)-STAT3. One representative experiment of three repeats is shown. ( h ) Phase-contrast microphotographs (PC) and immunofluorescence labelling of OCT4, NANOG, SSEA4 and TRA-1–81 in TL2i-OS3–10 cells after culturing in 2i/LIF medium without 4′-OHT for 30 passages. ( i ) Western blot analysis of STAT3 and STAT3-ER T2 expression in TL-OS3–10 cells and TL2i-OS3–10 cells (+/−4′-OHT), analysed with antibodies to total STAT3. One representative experiment of two repeats is shown. ( j ) Phase-contrast microphotographs (PC), AP detection and immunofluorescence labelling of OCT4, NANOG, TRA-1–81 and SSEA4 in TL2i-OS3–10 cells after culturing in N2B27+2i/LIF basal medium for eight passages. ( k ) Histogram representation of the cloning efficiency of F, TL and TL2i cells (OS3–10, H9S3–2 and H9S3–14 lines) after single-cell dissociation with 0.05% trypsin-EDTA and re-plating on feeders in the presence of 10 μM ROCK inhibitor Y-27632 for 24 h post dissociation. Tukey's test; n =3; error bars indicate the mean±s. e. Scale bar, 50 μm ( a , d , h , i ).

    Article Snippet: TL2i cells were regularly passaged by single-cell dissociation with 0.05% trypsin-EDTA (Gibco) every 4 days.

    Techniques: Activity Assay, Immunofluorescence, Staining, Western Blot, Expressing, Clone Assay