Immunofluorescence:Article Title: Spatiotemporal transcriptomic profiling reveals upregulation of glycolysis pathway genes before overt tauopathy in the PS19 mouse model.
Article Snippet: .. For immunofluorescence staining, the sections were incubated in Background Punisher (Biocare Medical) for 20 min, then primary antibodies (SYN antibody, Santa Cruz, SC-12737, 1:100 dilution and PGK1 antibody, Cell Signaling, 63536, 1:200 dilution) for 1 h, followed by the respective secondary antibodies conjugated to Alexa Fluor (Life Technologies, 1:500 dilution) for 1 h. Nuclei were stained with Hoechst and the slides were mounted with VECTASHIELD aqueous antifade mounting medium (Vector Laboratories). ..
Article Title: Spatiotemporal transcriptomic profiling reveals upregulation of glycolysis pathway genes before overt tauopathy in the PS19 mouse model
Article Snippet: .. For immunofluorescence staining, the sections were incubated in Background Punisher (Biocare Medical) for 20 min, then primary antibodies (SYN antibody, Santa Cruz, SC-12737, 1:100 dilution and PGK1 antibody, Cell Signaling, 63536, 1:200 dilution) for 1 h, followed by the respective secondary antibodies conjugated to Alexa Fluor (Life Technologies, 1:500 dilution) for 1 h. Nuclei were stained with Hoechst and the slides were mounted with VECTASHIELD aqueous antifade mounting medium (Vector Laboratories). ..
Staining:Article Title: Spatiotemporal transcriptomic profiling reveals upregulation of glycolysis pathway genes before overt tauopathy in the PS19 mouse model.
Article Snippet: .. For immunofluorescence staining, the sections were incubated in Background Punisher (Biocare Medical) for 20 min, then primary antibodies (SYN antibody, Santa Cruz, SC-12737, 1:100 dilution and PGK1 antibody, Cell Signaling, 63536, 1:200 dilution) for 1 h, followed by the respective secondary antibodies conjugated to Alexa Fluor (Life Technologies, 1:500 dilution) for 1 h. Nuclei were stained with Hoechst and the slides were mounted with VECTASHIELD aqueous antifade mounting medium (Vector Laboratories). ..
Article Title: Spatiotemporal transcriptomic profiling reveals upregulation of glycolysis pathway genes before overt tauopathy in the PS19 mouse model
Article Snippet: .. For immunofluorescence staining, the sections were incubated in Background Punisher (Biocare Medical) for 20 min, then primary antibodies (SYN antibody, Santa Cruz, SC-12737, 1:100 dilution and PGK1 antibody, Cell Signaling, 63536, 1:200 dilution) for 1 h, followed by the respective secondary antibodies conjugated to Alexa Fluor (Life Technologies, 1:500 dilution) for 1 h. Nuclei were stained with Hoechst and the slides were mounted with VECTASHIELD aqueous antifade mounting medium (Vector Laboratories). ..
Article Title: Physalin A Suppresses Human Oral Squamous Carcinoma Cell Migration and Invasion Through Inhibiting Grb2/Ras and MMP/uPA Signaling Pathways.
Article Snippet: The quantified proteins were electrophoretically separated by 8-12% SDSPAGE gel (based on different KDa), and were transferred to polyvinylidene difluoride (PVDF; Cat. No. IPVH00010; Merck Millipore, Burlington, MA, USA) membranes, which were blocked with 5% albumin. .. The membranes were then stained using primary antibodies [anti- plasminogen activator inhibitor 1 (PAI-1; sc-5297; Santa Cruz Biotechnology), -uPA (sc-59727; Santa Cruz Biotechnology), -E-cad (sc-374067; Santa Cruz Biotechnology), -α-tubulin (α-tub; sc-8035; Santa Cruz Biotechnology), -MMP-1 (sc21731; Santa Cruz Biotechnology), -MMP-2 (sc-13595; Santa Cruz Biotechnology), -MMP-3 (sc-21732; Santa Cruz Biotechnology), -MMP-9 (sc-393859; Santa Cruz Biotechnology), -tissue inhibitor of metalloprotease (TIMP1; sc-365905; Santa Cruz Biotechnology), -TIMP-2 (sc21735; Santa Cruz Biotechnology), -p-IKK (p-IKK; sc293135; Santa Cruz Biotechnology), -IKK (sc-8032; Santa Cruz Biotechnology), -p-IκB (sc-8404; Santa Cruz Biotechnology), -IκB (sc-1643; Santa Cruz Biotechnology), -NF-κB p65 (sc-8008; Santa Cruz Biotechnology), -NF-κB p50 (sc-8414; Santa Cruz Biotechnology), -p-ERK (9911; Cell Signaling Technology), -ERK (9911; Cell Signaling Technology), -p-c-Jun N-terminal kinase (-p-JNK; 4668; Cell Signaling Technology), -JNK (9252; Cell Signaling Technology), -p-p38 (9211; Cell Signaling Technology), -p38 (9212; Cell Signaling Technology), -p-c-Jun (3270; Cell Signaling Technology), -c-Jun (9165; Cell Signaling Technology), -Grb2 (3972; Cell Signaling Technology), -Ras (3965; Cell Signaling Technology), -p-PI3K (4228; Cell Signaling Technology), -PI3K (4292; Cell Signaling Technology), -p-Akt (9271; Cell Signaling Technology), -Akt (9272; Cell Signaling Technology), -PCNA (sc-56; Santa Cruz Biotechnology) and -β-actin (sc-47778; Santa Cruz Biotechnology)] in 4°C overnight. ..
Article Title: Physalin A Suppresses Human Oral Squamous Carcinoma Cell Migration and Invasion Through Inhibiting Grb2/Ras and MMP/uPA Signaling Pathways
Article Snippet: The quantified proteins were electrophoretically separated by 8-12% SDS-PAGE gel (based on different KDa), and were transferred to polyvinylidene difluoride (PVDF; Cat. No. IPVH00010; Merck Millipore, Burlington, MA, USA) membranes, which were blocked with 5% albumin. .. The membranes were then stained using primary antibodies [anti- plasminogen activator inhibitor 1 (PAI-1; sc-5297; Santa Cruz Biotechnology), -uPA (sc-59727; Santa Cruz Biotechnology), -E-cad (sc-374067; Santa Cruz Biotechnology), -α-tubulin (α-tub; sc-8035; Santa Cruz Biotechnology), -MMP-1 (sc-21731; Santa Cruz Biotechnology), -MMP-2 (sc-13595; Santa Cruz Biotechnology), -MMP-3 (sc-21732; Santa Cruz Biotechnology), -MMP-9 (sc-393859; Santa Cruz Biotechnology), -tissue inhibitor of metalloprotease (TIMP-1; sc-365905; Santa Cruz Biotechnology), -TIMP-2 (sc-21735; Santa Cruz Biotechnology), -p-IKK (p-IKK; sc-293135; Santa Cruz Biotechnology), -IKK (sc-8032; Santa Cruz Biotechnology), -p-IκB (sc-8404; Santa Cruz Biotechnology), -IκB (sc-1643; Santa Cruz Biotechnology), -NF-κB p65 (sc-8008; Santa Cruz Biotechnology), -NF-κB p50 (sc-8414; Santa Cruz Biotechnology), -p-ERK (9911; Cell Signaling Technology), -ERK (9911; Cell Signaling Technology), -p-c-Jun N-terminal kinase (-p-JNK; 4668; Cell Signaling Technology), -JNK (9252; Cell Signaling Technology), -p-p38 (9211; Cell Signaling Technology), -p38 (9212; Cell Signaling Technology), -p-c-Jun (3270; Cell Signaling Technology), -c-Jun (9165; Cell Signaling Technology), -Grb2 (3972; Cell Signaling Technology), -Ras (3965; Cell Signaling Technology), -p-PI3K (4228; Cell Signaling Technology), -PI3K (4292; Cell Signaling Technology), -p-Akt (9271; Cell Signaling Technology), -Akt (9272; Cell Signaling Technology), -PCNA (sc-56; Santa Cruz Biotechnology) and -β-actin (sc-47778; Santa Cruz Biotechnology)] in 4°C overnight. ..
Incubation:Article Title: Spatiotemporal transcriptomic profiling reveals upregulation of glycolysis pathway genes before overt tauopathy in the PS19 mouse model.
Article Snippet: .. For immunofluorescence staining, the sections were incubated in Background Punisher (Biocare Medical) for 20 min, then primary antibodies (SYN antibody, Santa Cruz, SC-12737, 1:100 dilution and PGK1 antibody, Cell Signaling, 63536, 1:200 dilution) for 1 h, followed by the respective secondary antibodies conjugated to Alexa Fluor (Life Technologies, 1:500 dilution) for 1 h. Nuclei were stained with Hoechst and the slides were mounted with VECTASHIELD aqueous antifade mounting medium (Vector Laboratories). ..
Article Title: Spatiotemporal transcriptomic profiling reveals upregulation of glycolysis pathway genes before overt tauopathy in the PS19 mouse model
Article Snippet: .. For immunofluorescence staining, the sections were incubated in Background Punisher (Biocare Medical) for 20 min, then primary antibodies (SYN antibody, Santa Cruz, SC-12737, 1:100 dilution and PGK1 antibody, Cell Signaling, 63536, 1:200 dilution) for 1 h, followed by the respective secondary antibodies conjugated to Alexa Fluor (Life Technologies, 1:500 dilution) for 1 h. Nuclei were stained with Hoechst and the slides were mounted with VECTASHIELD aqueous antifade mounting medium (Vector Laboratories). ..
Article Title: Molecular interactome of HNRNPU reveals regulatory networks in neuronal differentiation and DNA methylation.
Article Snippet: .. Primary antibodies [HNRNPU (Santa Cruz Biotechnology, sc-32315, 1:25), HNRNPU (Novus Bio, NBP2-49290, 1:500), FUS (Cell Signaling T echnology, 67840S, 1:1000), ARID1B (Cell Signaling T echnology, 92964S, 1:1000), EEF2 (Proteintech, 20107-1-AP, 1:500), NUFIP2 (Proteintech, 67195-1-Ig, 1:1000), DDX17 (Novus Bio, NB200-352, 1:500), and Fibrillarin (Cell Signaling Technology, 2639, 1:500)] were diluted in blocking buffer and incubated overnight at + 4 ◦C. .. After washing with 1 × DPBS, secondary antibodies [Anti-Mouse 555 (Biotium), and Anti-Rabbit 488 (Invitrogen)] diluted 1:1000 and Hoechst (1:2000, Invitrogen) in blocking buffer were incubated 1 h at room temperature, protected from light.
Article Title: Protective Effect of Silymarin Against Paclitaxel‐Induced Cardiotoxicity
Article Snippet: .. The membranes were blocked with 5% w/v bovine serum albumin for 90 min. Primary antibodies (IL‐1β [sc‐52,012], TNF‐α [sc‐52,746], Caspase‐3 [sc‐56,053], Bcl‐2 [sc‐7382], NF‐κB‐p65 [sc‐109], and beta‐actin [sc‐47,778], Santa Cruz Biotechnology, USA; P2X7R [11144–1‐AP], Proteintech Group, USA) were applied to the membranes and incubated at 4°C for approximately 15 h. Following a wash with Tris‐buffered saline containing Tween, the membranes were exposed to secondary antibodies (sc‐2004/sc‐2005, Santa Cruz Biotechnology, USA) conjugated with horseradish peroxidase (TP‐125‐HL, Thermo Fisher Scientific, USA) for 90 min. β‐actin (1:2000 dilution) was used as the loading control. .. Protein bands were visualized using enhanced Western ECL substrate (3405, Thermo Fisher Scientific, USA) and analyzed with software (Image Lab, Bio‐Rad, USA).
Binding Assay:Article Title: OsRALF26 Serves as an Endogenous Signal Recognised by XA21 to Promote Robust and Distal Resistance in Rice.
Article Snippet: Total proteins (150–250 μg per well) were separated on 8%–15% SDS–PAGE gels and transferred to PVDF or nitrocellulose membranes (BioRad). .. For detection, primary antibodies [anti- GFP (SC- 9996), anti- Myc (SC- 47694), anti- His (SC- 8036), and anti- GST (SC53909); Santa Cruz Biotechnology] and an HRP- conjugated secondary antibody [mouse IgG lambda binding protein (m- IgGλ BP; SC- 516132- CM; Santa Cruz Biotechnology)] were used. .. Immunoreactive bands were visualised using the SuperSignal West Pico Chemiluminescent Substrate (Thermo Fisher Scientific).
Blocking Assay:Article Title: Molecular interactome of HNRNPU reveals regulatory networks in neuronal differentiation and DNA methylation.
Article Snippet: .. Primary antibodies [HNRNPU (Santa Cruz Biotechnology, sc-32315, 1:25), HNRNPU (Novus Bio, NBP2-49290, 1:500), FUS (Cell Signaling T echnology, 67840S, 1:1000), ARID1B (Cell Signaling T echnology, 92964S, 1:1000), EEF2 (Proteintech, 20107-1-AP, 1:500), NUFIP2 (Proteintech, 67195-1-Ig, 1:1000), DDX17 (Novus Bio, NB200-352, 1:500), and Fibrillarin (Cell Signaling Technology, 2639, 1:500)] were diluted in blocking buffer and incubated overnight at + 4 ◦C. .. After washing with 1 × DPBS, secondary antibodies [Anti-Mouse 555 (Biotium), and Anti-Rabbit 488 (Invitrogen)] diluted 1:1000 and Hoechst (1:2000, Invitrogen) in blocking buffer were incubated 1 h at room temperature, protected from light.
Saline:Article Title: Protective Effect of Silymarin Against Paclitaxel‐Induced Cardiotoxicity
Article Snippet: .. The membranes were blocked with 5% w/v bovine serum albumin for 90 min. Primary antibodies (IL‐1β [sc‐52,012], TNF‐α [sc‐52,746], Caspase‐3 [sc‐56,053], Bcl‐2 [sc‐7382], NF‐κB‐p65 [sc‐109], and beta‐actin [sc‐47,778], Santa Cruz Biotechnology, USA; P2X7R [11144–1‐AP], Proteintech Group, USA) were applied to the membranes and incubated at 4°C for approximately 15 h. Following a wash with Tris‐buffered saline containing Tween, the membranes were exposed to secondary antibodies (sc‐2004/sc‐2005, Santa Cruz Biotechnology, USA) conjugated with horseradish peroxidase (TP‐125‐HL, Thermo Fisher Scientific, USA) for 90 min. β‐actin (1:2000 dilution) was used as the loading control. .. Protein bands were visualized using enhanced Western ECL substrate (3405, Thermo Fisher Scientific, USA) and analyzed with software (Image Lab, Bio‐Rad, USA).
Control:Article Title: Protective Effect of Silymarin Against Paclitaxel‐Induced Cardiotoxicity
Article Snippet: .. The membranes were blocked with 5% w/v bovine serum albumin for 90 min. Primary antibodies (IL‐1β [sc‐52,012], TNF‐α [sc‐52,746], Caspase‐3 [sc‐56,053], Bcl‐2 [sc‐7382], NF‐κB‐p65 [sc‐109], and beta‐actin [sc‐47,778], Santa Cruz Biotechnology, USA; P2X7R [11144–1‐AP], Proteintech Group, USA) were applied to the membranes and incubated at 4°C for approximately 15 h. Following a wash with Tris‐buffered saline containing Tween, the membranes were exposed to secondary antibodies (sc‐2004/sc‐2005, Santa Cruz Biotechnology, USA) conjugated with horseradish peroxidase (TP‐125‐HL, Thermo Fisher Scientific, USA) for 90 min. β‐actin (1:2000 dilution) was used as the loading control. .. Protein bands were visualized using enhanced Western ECL substrate (3405, Thermo Fisher Scientific, USA) and analyzed with software (Image Lab, Bio‐Rad, USA).
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