Molecular Weight:Article Title: Optogenetic cross-linking of the actin cytoskeleton using DARPins
Article Snippet: .. DARPins were separated from unreacted peptide and the buffer was replaced with phosphate buffer (48 mM K2HPO4, 4.5 mM KH2PO4, 14 mM NaH2PO4 in distilled water at pH 7.2) by ZebaSpin Desalting columns with a molecular weight cut-off at 7 kDa (Thermo Fisher, #78606). ..
Negative Control:Article Title: Fc region variant
Article Snippet: .. A sample prepared by adding a phosphate buffer (pH 7.4) (Gibco) instead of the immune complex was used as the negative control. .. The produced plasmids were introduced transiently into the HEK293H cell line derived from human embryonic kidney cancer cells (Invitrogen) or into FreeStyle293 cells (Invitrogen) for antibody expression.
Membrane:Article Title: Human 3D epithelioids enable continuous long-term clonal evolution studies across multiple epithelial tissues
Article Snippet: .. Membrane biopsies from human epithelioid cultures were obtained by using a 5 mm diameter biopsy punch, and fixed in a fixative solution containing 4% PFA (Sigma-Aldrich, P6148-500G) and 2.5% glutaraldehyde (EM grade distillation purified, Electron Microscopy Sciences, 16210) in 0.1 M phosphate buffer pH 7.4 (PB) (0.2 M Na 2 HPO 4 , Acros Organics, 448150010; 54 mM NaH 2 PO 4 , Acros Organics, 447760010; dissolved in ddH 2 O, pH 7.4) at 37°C for 15 minutes. .. Membrane biopsies from human epithelioid cultures were obtained by using a 5 mm diameter biopsy punch, and fixed in a fixative solution containing 4% PFA (Sigma-Aldrich, P6148-500G) and 2.5% glutaraldehyde (EM grade distillation purified, Electron Microscopy Sciences, 16210) in 0.1 M phosphate buffer pH 7.4 (PB) (0.2 M Na 2 HPO 4 , Acros Organics, 448150010; 54 mM NaH 2 PO 4 , Acros Organics, 447760010; dissolved in ddH 2 O, pH 7.4) at 37°C for 15 minutes.
Distillation:Article Title: Human 3D epithelioids enable continuous long-term clonal evolution studies across multiple epithelial tissues
Article Snippet: .. Membrane biopsies from human epithelioid cultures were obtained by using a 5 mm diameter biopsy punch, and fixed in a fixative solution containing 4% PFA (Sigma-Aldrich, P6148-500G) and 2.5% glutaraldehyde (EM grade distillation purified, Electron Microscopy Sciences, 16210) in 0.1 M phosphate buffer pH 7.4 (PB) (0.2 M Na 2 HPO 4 , Acros Organics, 448150010; 54 mM NaH 2 PO 4 , Acros Organics, 447760010; dissolved in ddH 2 O, pH 7.4) at 37°C for 15 minutes. .. Membrane biopsies from human epithelioid cultures were obtained by using a 5 mm diameter biopsy punch, and fixed in a fixative solution containing 4% PFA (Sigma-Aldrich, P6148-500G) and 2.5% glutaraldehyde (EM grade distillation purified, Electron Microscopy Sciences, 16210) in 0.1 M phosphate buffer pH 7.4 (PB) (0.2 M Na 2 HPO 4 , Acros Organics, 448150010; 54 mM NaH 2 PO 4 , Acros Organics, 447760010; dissolved in ddH 2 O, pH 7.4) at 37°C for 15 minutes.
Purification:Article Title: Human 3D epithelioids enable continuous long-term clonal evolution studies across multiple epithelial tissues
Article Snippet: .. Membrane biopsies from human epithelioid cultures were obtained by using a 5 mm diameter biopsy punch, and fixed in a fixative solution containing 4% PFA (Sigma-Aldrich, P6148-500G) and 2.5% glutaraldehyde (EM grade distillation purified, Electron Microscopy Sciences, 16210) in 0.1 M phosphate buffer pH 7.4 (PB) (0.2 M Na 2 HPO 4 , Acros Organics, 448150010; 54 mM NaH 2 PO 4 , Acros Organics, 447760010; dissolved in ddH 2 O, pH 7.4) at 37°C for 15 minutes. .. Membrane biopsies from human epithelioid cultures were obtained by using a 5 mm diameter biopsy punch, and fixed in a fixative solution containing 4% PFA (Sigma-Aldrich, P6148-500G) and 2.5% glutaraldehyde (EM grade distillation purified, Electron Microscopy Sciences, 16210) in 0.1 M phosphate buffer pH 7.4 (PB) (0.2 M Na 2 HPO 4 , Acros Organics, 448150010; 54 mM NaH 2 PO 4 , Acros Organics, 447760010; dissolved in ddH 2 O, pH 7.4) at 37°C for 15 minutes.
Electron Microscopy:Article Title: Human 3D epithelioids enable continuous long-term clonal evolution studies across multiple epithelial tissues
Article Snippet: .. Membrane biopsies from human epithelioid cultures were obtained by using a 5 mm diameter biopsy punch, and fixed in a fixative solution containing 4% PFA (Sigma-Aldrich, P6148-500G) and 2.5% glutaraldehyde (EM grade distillation purified, Electron Microscopy Sciences, 16210) in 0.1 M phosphate buffer pH 7.4 (PB) (0.2 M Na 2 HPO 4 , Acros Organics, 448150010; 54 mM NaH 2 PO 4 , Acros Organics, 447760010; dissolved in ddH 2 O, pH 7.4) at 37°C for 15 minutes. .. Membrane biopsies from human epithelioid cultures were obtained by using a 5 mm diameter biopsy punch, and fixed in a fixative solution containing 4% PFA (Sigma-Aldrich, P6148-500G) and 2.5% glutaraldehyde (EM grade distillation purified, Electron Microscopy Sciences, 16210) in 0.1 M phosphate buffer pH 7.4 (PB) (0.2 M Na 2 HPO 4 , Acros Organics, 448150010; 54 mM NaH 2 PO 4 , Acros Organics, 447760010; dissolved in ddH 2 O, pH 7.4) at 37°C for 15 minutes.
Concentration Assay:
Spectrophotometry:
Muscles:Article Title: Genetic Rescue of a Lethal Wasting Mutation ( Syb1 lew/lew ) by Neuron-Specific Expression of ECFP-Syb2
Article Snippet: .. Briefly, lumbrical or triangularis sterni muscles were fixed in 2% paraformaldehyde in 0.1 M phosphate buffer (pH 7.3) overnight at 4 ° C. Muscle samples were then incubated with Texas-Red conjugated α-bungarotoxin (α-bgt) (2 nM, Invitrogen, Carlsbad, California, USA) for 30 minutes, followed by incubation with primary antibodies diluted in antibody dilution buffer (500 mM NaCl, 0.01 M phosphate buffer, 3% BSA and 0.01% thimerosal) overnight at 4 ° C. The following primary antibodies were used: rabbit polyclonal anti-synaptotagmin2 (I735) , rabbit polyclonal anti-synaptobrevin1 (P938) and rabbit polyclonal anti-synaptobrevin2 (P939) (generous gifts from Dr. Thomas Südhof, Stanford University School of Medicine, Palo Alto, CA, USA). .. After extensive washes, muscle samples were incubated overnight at 4 ° C with Alexa Fluor 647-conjugated goat anti-rabbit IgG (1:600, Jackson ImmunoResearch Laboratories).
Article Title: Loss of ACTA1 leads to delayed γ-AChR / ε-AChR switch in skeletal muscle in mice
Article Snippet: .. Briefly, either diaphragm or triangularis sterni muscles from Acta1 −/− and their littermate controls ( Acta1 +/+ or Acta1 +/− ) at E18.5 or P5 were fixed in 2% paraformaldehyde in 0.1 M phosphate buffer (pH 7.3) overnight at 4 ° C. Samples were extensively washed with PBS and then incubated with Texas-Red conjugated α-bungarotoxin (α-bgt) (2 nM, Invitrogen, Carlsbad, California, USA) for 30 minutes at room temperature. .. Samples were then incubated with primary antibodies overnight at 4 ° C. The following polyclonal antibodies were used: neurofilament (NF150) (Chemicon, Temecula, CA, USA), synaptotagmin 2 (I735), syntaxin 1(I375) (generous gifts from Dr. Thomas Südhof, Stanford University School of Medicine, Palo Alto, CA, USA) , and acetylcholinesterase (AChE) (generous gifts from Dr. Palmer Taylor, UC San Diego, CA, USA).
Incubation:Article Title: Genetic Rescue of a Lethal Wasting Mutation ( Syb1 lew/lew ) by Neuron-Specific Expression of ECFP-Syb2
Article Snippet: .. Briefly, lumbrical or triangularis sterni muscles were fixed in 2% paraformaldehyde in 0.1 M phosphate buffer (pH 7.3) overnight at 4 ° C. Muscle samples were then incubated with Texas-Red conjugated α-bungarotoxin (α-bgt) (2 nM, Invitrogen, Carlsbad, California, USA) for 30 minutes, followed by incubation with primary antibodies diluted in antibody dilution buffer (500 mM NaCl, 0.01 M phosphate buffer, 3% BSA and 0.01% thimerosal) overnight at 4 ° C. The following primary antibodies were used: rabbit polyclonal anti-synaptotagmin2 (I735) , rabbit polyclonal anti-synaptobrevin1 (P938) and rabbit polyclonal anti-synaptobrevin2 (P939) (generous gifts from Dr. Thomas Südhof, Stanford University School of Medicine, Palo Alto, CA, USA). .. After extensive washes, muscle samples were incubated overnight at 4 ° C with Alexa Fluor 647-conjugated goat anti-rabbit IgG (1:600, Jackson ImmunoResearch Laboratories).
Article Title: Loss of ACTA1 leads to delayed γ-AChR / ε-AChR switch in skeletal muscle in mice
Article Snippet: .. Briefly, either diaphragm or triangularis sterni muscles from Acta1 −/− and their littermate controls ( Acta1 +/+ or Acta1 +/− ) at E18.5 or P5 were fixed in 2% paraformaldehyde in 0.1 M phosphate buffer (pH 7.3) overnight at 4 ° C. Samples were extensively washed with PBS and then incubated with Texas-Red conjugated α-bungarotoxin (α-bgt) (2 nM, Invitrogen, Carlsbad, California, USA) for 30 minutes at room temperature. .. Samples were then incubated with primary antibodies overnight at 4 ° C. The following polyclonal antibodies were used: neurofilament (NF150) (Chemicon, Temecula, CA, USA), synaptotagmin 2 (I735), syntaxin 1(I375) (generous gifts from Dr. Thomas Südhof, Stanford University School of Medicine, Palo Alto, CA, USA) , and acetylcholinesterase (AChE) (generous gifts from Dr. Palmer Taylor, UC San Diego, CA, USA).
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