Membrane:Article Title: Investigating the Effect and Mechanism of Protocatechuic Aldehyde on Vascular Dementia Based on Multi-Omics Approach
Article Snippet: .. The membrane was placed in 5% skim milk and blocked on a shaker for 1 h. The membrane was incubated with primary antibody, including vesicular glutamate Transporter 1 (vGLUT1; 1:1000, 47181, Cell Signaling Technology, Danvers, MA, USA), MCT1 (1:1000, 20139-1-AP, Proteintech, Chicago, IL, USA), MCT2 (1:500, 20355-1-AP, Proteintech, Chicago, IL, USA), MCT4 (1:2000, 22787-1-AP, Proteintech, Chicago, IL, USA) glucose transporter 1 (GLUT-1; 1:1000, 21829-1-AP, Proteintech, Chicago, IL, USA), hexokinase 2 (HK2; 1:5000, 22029-1-AP, Proteintech, Chicago, IL, USA), phosphofructokinase (PFK; 1:2000, 13389-1-AP, Proteintech, Chicago, IL, USA), pyruvate dehydrogenase (PDH; 1:1000, 2784, Cell Signaling Technology, Danvers, MA, USA), lactate dehydrogenase A (LDHA; 1:5000, 21799-1-AP, Proteintech, Chicago, IL, USA), and lactate dehydrogenase B (LDHB; 1:20,000, 14824-1-AP, Proteintech, Chicago, IL, USA) diluted in TBST at 4 °C overnight. .. After being washed thrice with TBST on a shaker, the membrane was incubated with secondary antibody (Goat Anti-Rabbit and Goat Anti-Mouse IgG H&L HRP; 1:10,000; ab6721, ab205719, Abcam, Cambridge, UK) diluted in TBST at room temperature for 1 h. After being washed three times with TBST on a shaker, the developer solution (32209, Thermo Fisher Scientific, Pittsburgh, PA, USA) was prepared at a 1:1 ratio, and the membrane was developed and photographed using a developer instrument (Tanon 5200, Tanon, Tokyo, Japan).
Article Title: Investigating the Effect and Mechanism of Protocatechuic Aldehyde on Vascular Dementia Based on Multi-Omics Approach.
Article Snippet: .. The membrane was placed in 5% skim milk and blocked on a shaker for 1 h. The membrane was incubated with primary antibody, including vesicular glutamate Transporter 1 (vGLUT1; 1:1000, 47181, Cell Signaling Technology, Danvers, MA, USA), MCT1 (1:1000, 20139-1-AP, Proteintech, Chicago, IL, USA), MCT2 (1:500, 20355-1-AP, Proteintech, Chicago, IL, USA), MCT4 (1:2000, 22787-1-AP, Proteintech, Chicago, IL, USA) glucose transporter 1 (GLUT-1; 1:1000, 21829-1-AP, Proteintech, Chicago, IL, USA), hexokinase 2 (HK2; 1:5000, 22029-1-AP, Proteintech, Chicago, IL, USA), phosphofructokinase (PFK; 1:2000, 13389-1-AP, Proteintech, Chicago, IL, USA), pyruvate dehydrogenase (PDH; 1:1000, 2784, Cell Signaling Technology, Danvers, MA, USA), lactate https://doi.org/10.3390/biom16030411 dehydrogenase A (LDHA; 1:5000, 21799-1-AP, Proteintech, Chicago, IL, USA), and lactate dehydrogenase B (LDHB; 1:20,000, 14824-1-AP, Proteintech, Chicago, IL, USA) diluted in TBST at 4 ◦C overnight. .. After being washed thrice with TBST on a shaker, the membrane was incubated with secondary antibody (Goat Anti-Rabbit and Goat Anti-Mouse IgG H&L HRP; 1:10,000; ab6721, ab205719, Abcam, Cambridge, UK) diluted in TBST at room temperature for 1 h. After being washed three times with TBST on a shaker, the developer solution (32209, Thermo Fisher Scientific, Pittsburgh, PA, USA) was prepared at a 1:1 ratio, and the membrane was developed and photographed using a developer instrument (Tanon 5200, Tanon, Tokyo, Japan).
Incubation:Article Title: Investigating the Effect and Mechanism of Protocatechuic Aldehyde on Vascular Dementia Based on Multi-Omics Approach
Article Snippet: .. The membrane was placed in 5% skim milk and blocked on a shaker for 1 h. The membrane was incubated with primary antibody, including vesicular glutamate Transporter 1 (vGLUT1; 1:1000, 47181, Cell Signaling Technology, Danvers, MA, USA), MCT1 (1:1000, 20139-1-AP, Proteintech, Chicago, IL, USA), MCT2 (1:500, 20355-1-AP, Proteintech, Chicago, IL, USA), MCT4 (1:2000, 22787-1-AP, Proteintech, Chicago, IL, USA) glucose transporter 1 (GLUT-1; 1:1000, 21829-1-AP, Proteintech, Chicago, IL, USA), hexokinase 2 (HK2; 1:5000, 22029-1-AP, Proteintech, Chicago, IL, USA), phosphofructokinase (PFK; 1:2000, 13389-1-AP, Proteintech, Chicago, IL, USA), pyruvate dehydrogenase (PDH; 1:1000, 2784, Cell Signaling Technology, Danvers, MA, USA), lactate dehydrogenase A (LDHA; 1:5000, 21799-1-AP, Proteintech, Chicago, IL, USA), and lactate dehydrogenase B (LDHB; 1:20,000, 14824-1-AP, Proteintech, Chicago, IL, USA) diluted in TBST at 4 °C overnight. .. After being washed thrice with TBST on a shaker, the membrane was incubated with secondary antibody (Goat Anti-Rabbit and Goat Anti-Mouse IgG H&L HRP; 1:10,000; ab6721, ab205719, Abcam, Cambridge, UK) diluted in TBST at room temperature for 1 h. After being washed three times with TBST on a shaker, the developer solution (32209, Thermo Fisher Scientific, Pittsburgh, PA, USA) was prepared at a 1:1 ratio, and the membrane was developed and photographed using a developer instrument (Tanon 5200, Tanon, Tokyo, Japan).
Article Title: Investigating the Effect and Mechanism of Protocatechuic Aldehyde on Vascular Dementia Based on Multi-Omics Approach.
Article Snippet: .. The membrane was placed in 5% skim milk and blocked on a shaker for 1 h. The membrane was incubated with primary antibody, including vesicular glutamate Transporter 1 (vGLUT1; 1:1000, 47181, Cell Signaling Technology, Danvers, MA, USA), MCT1 (1:1000, 20139-1-AP, Proteintech, Chicago, IL, USA), MCT2 (1:500, 20355-1-AP, Proteintech, Chicago, IL, USA), MCT4 (1:2000, 22787-1-AP, Proteintech, Chicago, IL, USA) glucose transporter 1 (GLUT-1; 1:1000, 21829-1-AP, Proteintech, Chicago, IL, USA), hexokinase 2 (HK2; 1:5000, 22029-1-AP, Proteintech, Chicago, IL, USA), phosphofructokinase (PFK; 1:2000, 13389-1-AP, Proteintech, Chicago, IL, USA), pyruvate dehydrogenase (PDH; 1:1000, 2784, Cell Signaling Technology, Danvers, MA, USA), lactate https://doi.org/10.3390/biom16030411 dehydrogenase A (LDHA; 1:5000, 21799-1-AP, Proteintech, Chicago, IL, USA), and lactate dehydrogenase B (LDHB; 1:20,000, 14824-1-AP, Proteintech, Chicago, IL, USA) diluted in TBST at 4 ◦C overnight. .. After being washed thrice with TBST on a shaker, the membrane was incubated with secondary antibody (Goat Anti-Rabbit and Goat Anti-Mouse IgG H&L HRP; 1:10,000; ab6721, ab205719, Abcam, Cambridge, UK) diluted in TBST at room temperature for 1 h. After being washed three times with TBST on a shaker, the developer solution (32209, Thermo Fisher Scientific, Pittsburgh, PA, USA) was prepared at a 1:1 ratio, and the membrane was developed and photographed using a developer instrument (Tanon 5200, Tanon, Tokyo, Japan).
Article Title: Investigating the Effect and Mechanism of Protocatechuic Aldehyde on Vascular Dementia Based on Multi-Omics Approach.
Article Snippet: .. The 300 μL blocking solution was added and blocked at 25 ◦C for 1 h. The slides were placed in primary antibodies, including monocarboxylate Transporter 1 [MCT1], MCT2 (PA5-72957, PA5-76603, Thermo Fisher Scientific, Pittsburgh, PA, USA), MCT4, GFAP, NeuN (22787-1-AP, 60190-1-Ig, 26975-1-AP, Proteintech, Chicago, IL, USA), DAPI (4083, Cell Signaling Technology, Danvers, MA, USA), dilution ratio 1:200, and incubated at 4 ◦C overnight. .. The fluorescent secondary antibodies (Goat Anti-Mouse IgG H&L Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L Alexa Fluor® 647, dilution ratio 1:200, ab150113, ab150079, Abcam, Cambridge, UK) were added and incubated at 25 ◦C for 1 h. Using a fluorescence microscope (CKX53, Olympus Corporation, Tokyo, Japan), results were documented following the addition of anti-fade mounting medium, which was applied after a light-protected PBS wash.
Article Title: Investigating the Effect and Mechanism of Protocatechuic Aldehyde on Vascular Dementia Based on Multi-Omics Approach
Article Snippet: .. The 300 μL blocking solution was added and blocked at 25 °C for 1 h. The slides were placed in primary antibodies, including monocarboxylate Transporter 1 [MCT1], MCT2 (PA5-72957, PA5-76603, Thermo Fisher Scientific, Pittsburgh, PA, USA), MCT4, GFAP, NeuN (22787-1-AP, 60190-1-Ig, 26975-1-AP, Proteintech, Chicago, IL, USA), DAPI (4083, Cell Signaling Technology, Danvers, MA, USA), dilution ratio 1:200, and incubated at 4 °C overnight. .. The fluorescent secondary antibodies (Goat Anti-Mouse IgG H&L Alexa Fluor ® 488, Goat Anti-Rabbit IgG H&L Alexa Fluor ® 647, dilution ratio 1:200, ab150113, ab150079, Abcam, Cambridge, UK) were added and incubated at 25 °C for 1 h. Using a fluorescence microscope (CKX53, Olympus Corporation, Tokyo, Japan), results were documented following the addition of anti-fade mounting medium, which was applied after a light-protected PBS wash.
Blocking Assay:Article Title: Investigating the Effect and Mechanism of Protocatechuic Aldehyde on Vascular Dementia Based on Multi-Omics Approach.
Article Snippet: .. The 300 μL blocking solution was added and blocked at 25 ◦C for 1 h. The slides were placed in primary antibodies, including monocarboxylate Transporter 1 [MCT1], MCT2 (PA5-72957, PA5-76603, Thermo Fisher Scientific, Pittsburgh, PA, USA), MCT4, GFAP, NeuN (22787-1-AP, 60190-1-Ig, 26975-1-AP, Proteintech, Chicago, IL, USA), DAPI (4083, Cell Signaling Technology, Danvers, MA, USA), dilution ratio 1:200, and incubated at 4 ◦C overnight. .. The fluorescent secondary antibodies (Goat Anti-Mouse IgG H&L Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L Alexa Fluor® 647, dilution ratio 1:200, ab150113, ab150079, Abcam, Cambridge, UK) were added and incubated at 25 ◦C for 1 h. Using a fluorescence microscope (CKX53, Olympus Corporation, Tokyo, Japan), results were documented following the addition of anti-fade mounting medium, which was applied after a light-protected PBS wash.
Article Title: Investigating the Effect and Mechanism of Protocatechuic Aldehyde on Vascular Dementia Based on Multi-Omics Approach
Article Snippet: .. The 300 μL blocking solution was added and blocked at 25 °C for 1 h. The slides were placed in primary antibodies, including monocarboxylate Transporter 1 [MCT1], MCT2 (PA5-72957, PA5-76603, Thermo Fisher Scientific, Pittsburgh, PA, USA), MCT4, GFAP, NeuN (22787-1-AP, 60190-1-Ig, 26975-1-AP, Proteintech, Chicago, IL, USA), DAPI (4083, Cell Signaling Technology, Danvers, MA, USA), dilution ratio 1:200, and incubated at 4 °C overnight. .. The fluorescent secondary antibodies (Goat Anti-Mouse IgG H&L Alexa Fluor ® 488, Goat Anti-Rabbit IgG H&L Alexa Fluor ® 647, dilution ratio 1:200, ab150113, ab150079, Abcam, Cambridge, UK) were added and incubated at 25 °C for 1 h. Using a fluorescence microscope (CKX53, Olympus Corporation, Tokyo, Japan), results were documented following the addition of anti-fade mounting medium, which was applied after a light-protected PBS wash.
other:
|
Proteintech is a verified supplier 