Western Blot:Article Title: Effects of enriched environment on the expression of β-amyloid and transport-related proteins LRP1 and RAGE in chronic sleep-deprived mice
Article Snippet: A total of 20 μg of each protein sample was used for 10% sodium dodecyl sulfate polyacrylamide gel electrophoresis followed by wet transfer onto PVDF membranes (Millipore). .. The samples were blocked for 20 min using QuickBlockTM Western blocking solution (P0252; Beyotime), washed 3 times in TBST, and then incubated with primary antibodies overnight at 4°C. .. The following primary antibodies were used: Rabbit anti-mouse LRP1 primary antibody (ab92544, 1:1,000; Abcam), Rabbit anti-mouse RAGE primary antibody (ab3611, 1:1,000; Abcam), and Rabbit anti-mouse GAPDH primary antibody (AP0063, 1:5,000; Bioworld).
Article Title: Midkine Prevents Calcification of Aortic Valve Interstitial Cells via Intercellular Crosstalk
Article Snippet: The protein samples were resolved by SDS-PAGE (4–20% gels) and then transferred to PVDF membranes using a protein transfer instrument (eBlotL1, GENSCRIPT). .. After 15 min of QuickBlockTM Western blocking solution (Beyotime, P0252) at room temperature, the membrane was incubated overnight with the primary antibody at 4°C for RUNX2(CST, 8486s), ALP (Zenbio,220,678), and GAPDH (Proteintech,60004-1-I). qRT-PCR assay using Cell Total RNA isolation kit (FOREGENE, RE-O3113) was utilized to extract mRNA. .. HiScript III RT SuperMix (Vazyme, R323-01) was used to perform reverse transcription of mRNA.
Article Title: Artesunate treats obesity in male mice and non-human primates through GDF15/GFRAL signalling axis
Article Snippet: The 10-μg protein samples were separated by 4–12% SDS–PAGE with MOPS-SDS running buffer and transferred to polyvinylidene difluoride membranes (Bio-Rad Laboratories). .. Membranes were blocked with 5 QuickBlockTM Western Blocking Solution (P0252, Beyotime Biotechnology) for 1 h and then incubated with primary antibodies prepared using Western Primary Antibody Diluent (P0256, Beyotime Biotechnology) at 4 °C overnight. .. Membranes were incubated with HRP-conjugated secondary antibodies prepared using QuickBlockTM Western Secondary Antibody Diluent (P0258, Beyotime Biotechnology), and incubated for 1 h at room temperature.
Blocking Assay:Article Title: Effects of enriched environment on the expression of β-amyloid and transport-related proteins LRP1 and RAGE in chronic sleep-deprived mice
Article Snippet: A total of 20 μg of each protein sample was used for 10% sodium dodecyl sulfate polyacrylamide gel electrophoresis followed by wet transfer onto PVDF membranes (Millipore). .. The samples were blocked for 20 min using QuickBlockTM Western blocking solution (P0252; Beyotime), washed 3 times in TBST, and then incubated with primary antibodies overnight at 4°C. .. The following primary antibodies were used: Rabbit anti-mouse LRP1 primary antibody (ab92544, 1:1,000; Abcam), Rabbit anti-mouse RAGE primary antibody (ab3611, 1:1,000; Abcam), and Rabbit anti-mouse GAPDH primary antibody (AP0063, 1:5,000; Bioworld).
Article Title: Midkine Prevents Calcification of Aortic Valve Interstitial Cells via Intercellular Crosstalk
Article Snippet: The protein samples were resolved by SDS-PAGE (4–20% gels) and then transferred to PVDF membranes using a protein transfer instrument (eBlotL1, GENSCRIPT). .. After 15 min of QuickBlockTM Western blocking solution (Beyotime, P0252) at room temperature, the membrane was incubated overnight with the primary antibody at 4°C for RUNX2(CST, 8486s), ALP (Zenbio,220,678), and GAPDH (Proteintech,60004-1-I). qRT-PCR assay using Cell Total RNA isolation kit (FOREGENE, RE-O3113) was utilized to extract mRNA. .. HiScript III RT SuperMix (Vazyme, R323-01) was used to perform reverse transcription of mRNA.
Article Title: Artesunate treats obesity in male mice and non-human primates through GDF15/GFRAL signalling axis
Article Snippet: The 10-μg protein samples were separated by 4–12% SDS–PAGE with MOPS-SDS running buffer and transferred to polyvinylidene difluoride membranes (Bio-Rad Laboratories). .. Membranes were blocked with 5 QuickBlockTM Western Blocking Solution (P0252, Beyotime Biotechnology) for 1 h and then incubated with primary antibodies prepared using Western Primary Antibody Diluent (P0256, Beyotime Biotechnology) at 4 °C overnight. .. Membranes were incubated with HRP-conjugated secondary antibodies prepared using QuickBlockTM Western Secondary Antibody Diluent (P0258, Beyotime Biotechnology), and incubated for 1 h at room temperature.
Incubation:Article Title: Effects of enriched environment on the expression of β-amyloid and transport-related proteins LRP1 and RAGE in chronic sleep-deprived mice
Article Snippet: A total of 20 μg of each protein sample was used for 10% sodium dodecyl sulfate polyacrylamide gel electrophoresis followed by wet transfer onto PVDF membranes (Millipore). .. The samples were blocked for 20 min using QuickBlockTM Western blocking solution (P0252; Beyotime), washed 3 times in TBST, and then incubated with primary antibodies overnight at 4°C. .. The following primary antibodies were used: Rabbit anti-mouse LRP1 primary antibody (ab92544, 1:1,000; Abcam), Rabbit anti-mouse RAGE primary antibody (ab3611, 1:1,000; Abcam), and Rabbit anti-mouse GAPDH primary antibody (AP0063, 1:5,000; Bioworld).
Article Title: Midkine Prevents Calcification of Aortic Valve Interstitial Cells via Intercellular Crosstalk
Article Snippet: The protein samples were resolved by SDS-PAGE (4–20% gels) and then transferred to PVDF membranes using a protein transfer instrument (eBlotL1, GENSCRIPT). .. After 15 min of QuickBlockTM Western blocking solution (Beyotime, P0252) at room temperature, the membrane was incubated overnight with the primary antibody at 4°C for RUNX2(CST, 8486s), ALP (Zenbio,220,678), and GAPDH (Proteintech,60004-1-I). qRT-PCR assay using Cell Total RNA isolation kit (FOREGENE, RE-O3113) was utilized to extract mRNA. .. HiScript III RT SuperMix (Vazyme, R323-01) was used to perform reverse transcription of mRNA.
Article Title: Artesunate treats obesity in male mice and non-human primates through GDF15/GFRAL signalling axis
Article Snippet: The 10-μg protein samples were separated by 4–12% SDS–PAGE with MOPS-SDS running buffer and transferred to polyvinylidene difluoride membranes (Bio-Rad Laboratories). .. Membranes were blocked with 5 QuickBlockTM Western Blocking Solution (P0252, Beyotime Biotechnology) for 1 h and then incubated with primary antibodies prepared using Western Primary Antibody Diluent (P0256, Beyotime Biotechnology) at 4 °C overnight. .. Membranes were incubated with HRP-conjugated secondary antibodies prepared using QuickBlockTM Western Secondary Antibody Diluent (P0258, Beyotime Biotechnology), and incubated for 1 h at room temperature.
Membrane:Article Title: Midkine Prevents Calcification of Aortic Valve Interstitial Cells via Intercellular Crosstalk
Article Snippet: The protein samples were resolved by SDS-PAGE (4–20% gels) and then transferred to PVDF membranes using a protein transfer instrument (eBlotL1, GENSCRIPT). .. After 15 min of QuickBlockTM Western blocking solution (Beyotime, P0252) at room temperature, the membrane was incubated overnight with the primary antibody at 4°C for RUNX2(CST, 8486s), ALP (Zenbio,220,678), and GAPDH (Proteintech,60004-1-I). qRT-PCR assay using Cell Total RNA isolation kit (FOREGENE, RE-O3113) was utilized to extract mRNA. .. HiScript III RT SuperMix (Vazyme, R323-01) was used to perform reverse transcription of mRNA.
Article Title: Exploring the correlation between magnetic resonance diffusion tensor imaging (DTI) parameters and aquaporin expression and biochemical composition content in degenerative intervertebral disc nucleus pulposus tissue: a clinical experimental study.
Article Snippet: .. The following reagents were utilized in the study: AQP1 antibody (bs-1506R, Bioss, China), AQP3 antibody (bs-1253R, Bioss, China), GAPDH antibody (ab181602, Abcam, USA), goat anti-rabbit IgG secondary antibody (ab205718, Abcam, USA), Western primary antibody dilution (P0023A, Beyotime, China), QuickBlockTM Western blocking solution (P0252, Beyotime, China), Western rapid membrane transfer solution (10 ×) (P0572, Beyotime, China), SDS-PAGE protein loading buffer (5 ×, odorless) (P0286, Beyotime, China), BCA protein quantification kit (P0010S, Beyotime, China), One Step PAGE Gel Fast Preparation Kit (E303-01, Vazyme, China), Enhanced ECL Electrochemiluminescence Reagent (BL520A, Biosharp, China), Tissue Glycosaminoglycan (GAG) Total Content Dimethyl Methylene Blue (DMMB) Colorimetric Quantitative Detection Reagent (JM7861, Yuduobio, China), Sircol soluble collagen assay kit (S1000, Biocolor, UK), glacial acetic acid (A801296, Macklin, China), gastric protease (p6887, Sigma-Aldrich, Germany). .. Instruments used included a three-dimensional frozen grinding instrument (JXCL-3 K, Jingxin, China), constant temperature water bath (TSGP05, Thermo, USA), highspeed low-temperature centrifuge (5424R, Eppendorf, Germany), vertical electrophoresis instrument (PowerPac Basic, Bio-Rad, USA), universal protein transfer system (Trans-Blot Turbo, Bio-Rad, USA), automatic full-wavelength enzyme standard instrument (Infinite M200Pro, Tecan, Switzerland), automatic chemiluminescence image analysis system (5200, Tanon, China), and a 3.0T magnetic resonance imager (MR750, GE Discovery, USA).
Quantitative RT-PCR:Article Title: Midkine Prevents Calcification of Aortic Valve Interstitial Cells via Intercellular Crosstalk
Article Snippet: The protein samples were resolved by SDS-PAGE (4–20% gels) and then transferred to PVDF membranes using a protein transfer instrument (eBlotL1, GENSCRIPT). .. After 15 min of QuickBlockTM Western blocking solution (Beyotime, P0252) at room temperature, the membrane was incubated overnight with the primary antibody at 4°C for RUNX2(CST, 8486s), ALP (Zenbio,220,678), and GAPDH (Proteintech,60004-1-I). qRT-PCR assay using Cell Total RNA isolation kit (FOREGENE, RE-O3113) was utilized to extract mRNA. .. HiScript III RT SuperMix (Vazyme, R323-01) was used to perform reverse transcription of mRNA.
Isolation:Article Title: Midkine Prevents Calcification of Aortic Valve Interstitial Cells via Intercellular Crosstalk
Article Snippet: The protein samples were resolved by SDS-PAGE (4–20% gels) and then transferred to PVDF membranes using a protein transfer instrument (eBlotL1, GENSCRIPT). .. After 15 min of QuickBlockTM Western blocking solution (Beyotime, P0252) at room temperature, the membrane was incubated overnight with the primary antibody at 4°C for RUNX2(CST, 8486s), ALP (Zenbio,220,678), and GAPDH (Proteintech,60004-1-I). qRT-PCR assay using Cell Total RNA isolation kit (FOREGENE, RE-O3113) was utilized to extract mRNA. .. HiScript III RT SuperMix (Vazyme, R323-01) was used to perform reverse transcription of mRNA.
SDS Page:Article Title: Exploring the correlation between magnetic resonance diffusion tensor imaging (DTI) parameters and aquaporin expression and biochemical composition content in degenerative intervertebral disc nucleus pulposus tissue: a clinical experimental study.
Article Snippet: .. The following reagents were utilized in the study: AQP1 antibody (bs-1506R, Bioss, China), AQP3 antibody (bs-1253R, Bioss, China), GAPDH antibody (ab181602, Abcam, USA), goat anti-rabbit IgG secondary antibody (ab205718, Abcam, USA), Western primary antibody dilution (P0023A, Beyotime, China), QuickBlockTM Western blocking solution (P0252, Beyotime, China), Western rapid membrane transfer solution (10 ×) (P0572, Beyotime, China), SDS-PAGE protein loading buffer (5 ×, odorless) (P0286, Beyotime, China), BCA protein quantification kit (P0010S, Beyotime, China), One Step PAGE Gel Fast Preparation Kit (E303-01, Vazyme, China), Enhanced ECL Electrochemiluminescence Reagent (BL520A, Biosharp, China), Tissue Glycosaminoglycan (GAG) Total Content Dimethyl Methylene Blue (DMMB) Colorimetric Quantitative Detection Reagent (JM7861, Yuduobio, China), Sircol soluble collagen assay kit (S1000, Biocolor, UK), glacial acetic acid (A801296, Macklin, China), gastric protease (p6887, Sigma-Aldrich, Germany). .. Instruments used included a three-dimensional frozen grinding instrument (JXCL-3 K, Jingxin, China), constant temperature water bath (TSGP05, Thermo, USA), highspeed low-temperature centrifuge (5424R, Eppendorf, Germany), vertical electrophoresis instrument (PowerPac Basic, Bio-Rad, USA), universal protein transfer system (Trans-Blot Turbo, Bio-Rad, USA), automatic full-wavelength enzyme standard instrument (Infinite M200Pro, Tecan, Switzerland), automatic chemiluminescence image analysis system (5200, Tanon, China), and a 3.0T magnetic resonance imager (MR750, GE Discovery, USA).
Polyacrylamide Gel Electrophoresis:Article Title: Exploring the correlation between magnetic resonance diffusion tensor imaging (DTI) parameters and aquaporin expression and biochemical composition content in degenerative intervertebral disc nucleus pulposus tissue: a clinical experimental study.
Article Snippet: .. The following reagents were utilized in the study: AQP1 antibody (bs-1506R, Bioss, China), AQP3 antibody (bs-1253R, Bioss, China), GAPDH antibody (ab181602, Abcam, USA), goat anti-rabbit IgG secondary antibody (ab205718, Abcam, USA), Western primary antibody dilution (P0023A, Beyotime, China), QuickBlockTM Western blocking solution (P0252, Beyotime, China), Western rapid membrane transfer solution (10 ×) (P0572, Beyotime, China), SDS-PAGE protein loading buffer (5 ×, odorless) (P0286, Beyotime, China), BCA protein quantification kit (P0010S, Beyotime, China), One Step PAGE Gel Fast Preparation Kit (E303-01, Vazyme, China), Enhanced ECL Electrochemiluminescence Reagent (BL520A, Biosharp, China), Tissue Glycosaminoglycan (GAG) Total Content Dimethyl Methylene Blue (DMMB) Colorimetric Quantitative Detection Reagent (JM7861, Yuduobio, China), Sircol soluble collagen assay kit (S1000, Biocolor, UK), glacial acetic acid (A801296, Macklin, China), gastric protease (p6887, Sigma-Aldrich, Germany). .. Instruments used included a three-dimensional frozen grinding instrument (JXCL-3 K, Jingxin, China), constant temperature water bath (TSGP05, Thermo, USA), highspeed low-temperature centrifuge (5424R, Eppendorf, Germany), vertical electrophoresis instrument (PowerPac Basic, Bio-Rad, USA), universal protein transfer system (Trans-Blot Turbo, Bio-Rad, USA), automatic full-wavelength enzyme standard instrument (Infinite M200Pro, Tecan, Switzerland), automatic chemiluminescence image analysis system (5200, Tanon, China), and a 3.0T magnetic resonance imager (MR750, GE Discovery, USA).
Electrochemiluminescence:Article Title: Exploring the correlation between magnetic resonance diffusion tensor imaging (DTI) parameters and aquaporin expression and biochemical composition content in degenerative intervertebral disc nucleus pulposus tissue: a clinical experimental study.
Article Snippet: .. The following reagents were utilized in the study: AQP1 antibody (bs-1506R, Bioss, China), AQP3 antibody (bs-1253R, Bioss, China), GAPDH antibody (ab181602, Abcam, USA), goat anti-rabbit IgG secondary antibody (ab205718, Abcam, USA), Western primary antibody dilution (P0023A, Beyotime, China), QuickBlockTM Western blocking solution (P0252, Beyotime, China), Western rapid membrane transfer solution (10 ×) (P0572, Beyotime, China), SDS-PAGE protein loading buffer (5 ×, odorless) (P0286, Beyotime, China), BCA protein quantification kit (P0010S, Beyotime, China), One Step PAGE Gel Fast Preparation Kit (E303-01, Vazyme, China), Enhanced ECL Electrochemiluminescence Reagent (BL520A, Biosharp, China), Tissue Glycosaminoglycan (GAG) Total Content Dimethyl Methylene Blue (DMMB) Colorimetric Quantitative Detection Reagent (JM7861, Yuduobio, China), Sircol soluble collagen assay kit (S1000, Biocolor, UK), glacial acetic acid (A801296, Macklin, China), gastric protease (p6887, Sigma-Aldrich, Germany). .. Instruments used included a three-dimensional frozen grinding instrument (JXCL-3 K, Jingxin, China), constant temperature water bath (TSGP05, Thermo, USA), highspeed low-temperature centrifuge (5424R, Eppendorf, Germany), vertical electrophoresis instrument (PowerPac Basic, Bio-Rad, USA), universal protein transfer system (Trans-Blot Turbo, Bio-Rad, USA), automatic full-wavelength enzyme standard instrument (Infinite M200Pro, Tecan, Switzerland), automatic chemiluminescence image analysis system (5200, Tanon, China), and a 3.0T magnetic resonance imager (MR750, GE Discovery, USA).
Collagen Assay:Article Title: Exploring the correlation between magnetic resonance diffusion tensor imaging (DTI) parameters and aquaporin expression and biochemical composition content in degenerative intervertebral disc nucleus pulposus tissue: a clinical experimental study.
Article Snippet: .. The following reagents were utilized in the study: AQP1 antibody (bs-1506R, Bioss, China), AQP3 antibody (bs-1253R, Bioss, China), GAPDH antibody (ab181602, Abcam, USA), goat anti-rabbit IgG secondary antibody (ab205718, Abcam, USA), Western primary antibody dilution (P0023A, Beyotime, China), QuickBlockTM Western blocking solution (P0252, Beyotime, China), Western rapid membrane transfer solution (10 ×) (P0572, Beyotime, China), SDS-PAGE protein loading buffer (5 ×, odorless) (P0286, Beyotime, China), BCA protein quantification kit (P0010S, Beyotime, China), One Step PAGE Gel Fast Preparation Kit (E303-01, Vazyme, China), Enhanced ECL Electrochemiluminescence Reagent (BL520A, Biosharp, China), Tissue Glycosaminoglycan (GAG) Total Content Dimethyl Methylene Blue (DMMB) Colorimetric Quantitative Detection Reagent (JM7861, Yuduobio, China), Sircol soluble collagen assay kit (S1000, Biocolor, UK), glacial acetic acid (A801296, Macklin, China), gastric protease (p6887, Sigma-Aldrich, Germany). .. Instruments used included a three-dimensional frozen grinding instrument (JXCL-3 K, Jingxin, China), constant temperature water bath (TSGP05, Thermo, USA), highspeed low-temperature centrifuge (5424R, Eppendorf, Germany), vertical electrophoresis instrument (PowerPac Basic, Bio-Rad, USA), universal protein transfer system (Trans-Blot Turbo, Bio-Rad, USA), automatic full-wavelength enzyme standard instrument (Infinite M200Pro, Tecan, Switzerland), automatic chemiluminescence image analysis system (5200, Tanon, China), and a 3.0T magnetic resonance imager (MR750, GE Discovery, USA).
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