Incubation:Article Title: Hypothalamic SLC7A14 accounts for aging-reduced lipolysis in white adipose tissue of male mice.
Article Snippet: .. Addition of anti-mTOR antibody (1:100, Cat. No. 66888-1-Ig, Proteintech) into each sample and incubated with gentle rotation for 2 h. Then protein G-sepharose was added and incubated for 2 h. Immunoprecipitates were washed 4 times, followed by adding 45 uL kinase buffer (25mMHEPES pH 7.5, 100mMpotassium acetate, 1mMMgCl2) containing 500 uMATP (Cat. No. A600020-0005, sangon, Shanghai, China) and 500ng inactive S6K (Cat. No. P02664, Solarbio) to each sample and incubated at 37 °C for 30min, terminated by the addition of 2 x SDS sample buffer and analyzed by immunoblotting. .. The metabolite assessment of samples was performed by untargeted metabolomics at Novogene Biotech Co., Ltd. Serumwas collected from control or overexpressionof SLC7A14 inPOMCneuronmice.
Article Title: Hypothalamic SLC7A14 accounts for aging-reduced lipolysis in white adipose tissue of male mice
Article Snippet: .. Addition of anti-mTOR antibody (1:100, Cat. No. 66888-1-Ig, Proteintech) into each sample and incubated with gentle rotation for 2 h. Then protein G-sepharose was added and incubated for 2 h. Immunoprecipitates were washed 4 times, followed by adding 45 uL kinase buffer (25 mM HEPES pH 7.5, 100 mM potassium acetate, 1 mM MgCl 2 ) containing 500 uM ATP (Cat. No. A600020-0005, sangon, Shanghai, China) and 500 ng inactive S6K (Cat. No. P02664, Solarbio) to each sample and incubated at 37 °C for 30 min, terminated by the addition of 2 x SDS sample buffer and analyzed by immunoblotting. .. The metabolite assessment of samples was performed by untargeted metabolomics at Novogene Biotech Co., Ltd. Serum was collected from control or overexpression of SLC7A14 in POMC neuron mice.
Article Title: Metabolomics and proteomics reveal blocking argininosuccinate synthetase 1 alleviates colitis in mice
Article Snippet: .. The membrane was incubated with the following primary antibodies: anti-ASS1 antibody (16210-1-AP, Proteintech), anti-COX-2 antibody (12282, CST), anti-S6 antibody (2317, CST), anti-p-S6 antibody (4858, CST), anti-STAT3 antibody (4904, CST), anti-p-STAT3 antibody (9134, CST), anti-mTOR antibody (66889-1-Ig, Proteintech), anti-p-mTOR antibody (5536, CST), anti-ASL antibody (16645-1-AP, Proteintech), anti-IL1β antibody (66737-1-Ig, Proteintech) anti-GAPDH monoclonal antibody (HRP-60004, Proteintech), HRP-linked anti-rabbit IgG (SA00001-2, Proteintech), HRP-linked anti-mouse IgG (SA00001-1, Proteintech). .. Semi-quantitative analysis of each protein was performed using ImageJ software (version 1.5), and the band densities were normalized to the band density of GAPDH.
Article Title: Interfacial Coordination‐Engineered Emulgel Modulates the Osteo‐Immune Microenvironment in Periodontitis Treatment Via HIF1 Signaling Pathway‐Induced Metabolic Reprogramming
Article Snippet: .. Antibodies used in western blot as follows: mouse anti‐MYD88 antibody (1:2000, 67969‐1‐Ig, Proteintech, China), mouse anti‐mTOR antibody (1:25 000, 66888‐1‐Ig, Proteintech, China), rabbit anti‐AMPK antibody, rabbit anti‐HIF‐1α(1:1000, ab207442, Abcam, USA), mouse anti‐NF‐kB (1:1000, # Q04206 , Cell Signaling Technology, USA), mouse anti‐TNF‐α (Proteintech, 60291‐1‐Ig, 1:1000), rabbit anti‐iNOS (1:200, 22226‐1‐AP, Proteintech, China), mouse anti‐NLRP3 (1:2000, 68102‐1‐lg, Proteintech, China), rabbit anti‐IL‐1β (1:1000, ab254360, Abcam), mouse anti‐Tubulin (1:100, sc‐5286, Santa Cruz Biotechnology, USA), IRF4 (1:1000, ab315394, Abcam, USA), mouse anti‐STAT3 (1:1000, #9139, Cell Signaling Technology, USA), rabbit anti‐GLUT1(1:1000, 21829‐1‐AP, Proteintech, China), rabbit anti‐ERK1/2 (1:500, WL01864, Wanleibio), rabbit anti‐p‐ERK1/2 (1:1000, 28733‐1‐AP, Proteintech, China), mouse anti‐AKT(1:100, sc‐5298, Santa Cruz Biotechnology, USA), rabbit anti‐RUNX2(1:200, 20700‐1‐AP, Proteintech, China), and then incubated with HRP (horseradish peroxidase) conjugated secondary antibodies, anti‐rabbit (1:5000, Cat No. SA00001‐2, Proteintech, China) and anti‐mouse (1:5000, Cat No. SA00001‐1, Proteintech, China). .. The bound antibody was detected using a standard chemiluminescence method with the ChemiDoc Touch Imaging System. (Bio‐Rad, Universal Hood ll, USA).
Article Title: Metabolomics and proteomics reveal blocking argininosuccinate synthetase 1 alleviates colitis in mice.
Article Snippet: .. The membrane was incubated with the following primary antibodies: anti-ASS1 antibody (16210-1-AP, Proteintech), anti-COX-2 antibody (12282, CST), anti-S6 antibody (2317, CST), anti-p-S6 antibody (4858, CST), anti-STAT3 antibody (4904, CST), anti-p-STAT3 antibody (9134, CST), anti-mTOR antibody (66889- 1-Ig, Proteintech), anti-p-mTOR antibody (5536, CST), anti-ASL antibody (16645-1-AP, Proteintech), anti-IL1β antibody (66737-1-Ig, Proteintech) anti-GAPDH monoclonal antibody (HRP-60004, Proteintech), HRP-linked anti-rabbit IgG (SA00001-2, Proteintech), HRP-linked anti-mouse IgG (SA00001-1, Proteintech). .. Semiquantitative analysis of each protein was performed using ImageJ software (version 1.5), and the band densities were normalized to the band density of GAPDH.
Gentle:Article Title: Hypothalamic SLC7A14 accounts for aging-reduced lipolysis in white adipose tissue of male mice.
Article Snippet: .. Addition of anti-mTOR antibody (1:100, Cat. No. 66888-1-Ig, Proteintech) into each sample and incubated with gentle rotation for 2 h. Then protein G-sepharose was added and incubated for 2 h. Immunoprecipitates were washed 4 times, followed by adding 45 uL kinase buffer (25mMHEPES pH 7.5, 100mMpotassium acetate, 1mMMgCl2) containing 500 uMATP (Cat. No. A600020-0005, sangon, Shanghai, China) and 500ng inactive S6K (Cat. No. P02664, Solarbio) to each sample and incubated at 37 °C for 30min, terminated by the addition of 2 x SDS sample buffer and analyzed by immunoblotting. .. The metabolite assessment of samples was performed by untargeted metabolomics at Novogene Biotech Co., Ltd. Serumwas collected from control or overexpressionof SLC7A14 inPOMCneuronmice.
Article Title: Hypothalamic SLC7A14 accounts for aging-reduced lipolysis in white adipose tissue of male mice
Article Snippet: .. Addition of anti-mTOR antibody (1:100, Cat. No. 66888-1-Ig, Proteintech) into each sample and incubated with gentle rotation for 2 h. Then protein G-sepharose was added and incubated for 2 h. Immunoprecipitates were washed 4 times, followed by adding 45 uL kinase buffer (25 mM HEPES pH 7.5, 100 mM potassium acetate, 1 mM MgCl 2 ) containing 500 uM ATP (Cat. No. A600020-0005, sangon, Shanghai, China) and 500 ng inactive S6K (Cat. No. P02664, Solarbio) to each sample and incubated at 37 °C for 30 min, terminated by the addition of 2 x SDS sample buffer and analyzed by immunoblotting. .. The metabolite assessment of samples was performed by untargeted metabolomics at Novogene Biotech Co., Ltd. Serum was collected from control or overexpression of SLC7A14 in POMC neuron mice.
Western Blot:Article Title: Hypothalamic SLC7A14 accounts for aging-reduced lipolysis in white adipose tissue of male mice.
Article Snippet: .. Addition of anti-mTOR antibody (1:100, Cat. No. 66888-1-Ig, Proteintech) into each sample and incubated with gentle rotation for 2 h. Then protein G-sepharose was added and incubated for 2 h. Immunoprecipitates were washed 4 times, followed by adding 45 uL kinase buffer (25mMHEPES pH 7.5, 100mMpotassium acetate, 1mMMgCl2) containing 500 uMATP (Cat. No. A600020-0005, sangon, Shanghai, China) and 500ng inactive S6K (Cat. No. P02664, Solarbio) to each sample and incubated at 37 °C for 30min, terminated by the addition of 2 x SDS sample buffer and analyzed by immunoblotting. .. The metabolite assessment of samples was performed by untargeted metabolomics at Novogene Biotech Co., Ltd. Serumwas collected from control or overexpressionof SLC7A14 inPOMCneuronmice.
Article Title: Hypothalamic SLC7A14 accounts for aging-reduced lipolysis in white adipose tissue of male mice
Article Snippet: .. Addition of anti-mTOR antibody (1:100, Cat. No. 66888-1-Ig, Proteintech) into each sample and incubated with gentle rotation for 2 h. Then protein G-sepharose was added and incubated for 2 h. Immunoprecipitates were washed 4 times, followed by adding 45 uL kinase buffer (25 mM HEPES pH 7.5, 100 mM potassium acetate, 1 mM MgCl 2 ) containing 500 uM ATP (Cat. No. A600020-0005, sangon, Shanghai, China) and 500 ng inactive S6K (Cat. No. P02664, Solarbio) to each sample and incubated at 37 °C for 30 min, terminated by the addition of 2 x SDS sample buffer and analyzed by immunoblotting. .. The metabolite assessment of samples was performed by untargeted metabolomics at Novogene Biotech Co., Ltd. Serum was collected from control or overexpression of SLC7A14 in POMC neuron mice.
Article Title: Interfacial Coordination‐Engineered Emulgel Modulates the Osteo‐Immune Microenvironment in Periodontitis Treatment Via HIF1 Signaling Pathway‐Induced Metabolic Reprogramming
Article Snippet: .. Antibodies used in western blot as follows: mouse anti‐MYD88 antibody (1:2000, 67969‐1‐Ig, Proteintech, China), mouse anti‐mTOR antibody (1:25 000, 66888‐1‐Ig, Proteintech, China), rabbit anti‐AMPK antibody, rabbit anti‐HIF‐1α(1:1000, ab207442, Abcam, USA), mouse anti‐NF‐kB (1:1000, # Q04206 , Cell Signaling Technology, USA), mouse anti‐TNF‐α (Proteintech, 60291‐1‐Ig, 1:1000), rabbit anti‐iNOS (1:200, 22226‐1‐AP, Proteintech, China), mouse anti‐NLRP3 (1:2000, 68102‐1‐lg, Proteintech, China), rabbit anti‐IL‐1β (1:1000, ab254360, Abcam), mouse anti‐Tubulin (1:100, sc‐5286, Santa Cruz Biotechnology, USA), IRF4 (1:1000, ab315394, Abcam, USA), mouse anti‐STAT3 (1:1000, #9139, Cell Signaling Technology, USA), rabbit anti‐GLUT1(1:1000, 21829‐1‐AP, Proteintech, China), rabbit anti‐ERK1/2 (1:500, WL01864, Wanleibio), rabbit anti‐p‐ERK1/2 (1:1000, 28733‐1‐AP, Proteintech, China), mouse anti‐AKT(1:100, sc‐5298, Santa Cruz Biotechnology, USA), rabbit anti‐RUNX2(1:200, 20700‐1‐AP, Proteintech, China), and then incubated with HRP (horseradish peroxidase) conjugated secondary antibodies, anti‐rabbit (1:5000, Cat No. SA00001‐2, Proteintech, China) and anti‐mouse (1:5000, Cat No. SA00001‐1, Proteintech, China). .. The bound antibody was detected using a standard chemiluminescence method with the ChemiDoc Touch Imaging System. (Bio‐Rad, Universal Hood ll, USA).
Membrane:Article Title: Metabolomics and proteomics reveal blocking argininosuccinate synthetase 1 alleviates colitis in mice
Article Snippet: .. The membrane was incubated with the following primary antibodies: anti-ASS1 antibody (16210-1-AP, Proteintech), anti-COX-2 antibody (12282, CST), anti-S6 antibody (2317, CST), anti-p-S6 antibody (4858, CST), anti-STAT3 antibody (4904, CST), anti-p-STAT3 antibody (9134, CST), anti-mTOR antibody (66889-1-Ig, Proteintech), anti-p-mTOR antibody (5536, CST), anti-ASL antibody (16645-1-AP, Proteintech), anti-IL1β antibody (66737-1-Ig, Proteintech) anti-GAPDH monoclonal antibody (HRP-60004, Proteintech), HRP-linked anti-rabbit IgG (SA00001-2, Proteintech), HRP-linked anti-mouse IgG (SA00001-1, Proteintech). .. Semi-quantitative analysis of each protein was performed using ImageJ software (version 1.5), and the band densities were normalized to the band density of GAPDH.
Article Title: Metabolomics and proteomics reveal blocking argininosuccinate synthetase 1 alleviates colitis in mice.
Article Snippet: .. The membrane was incubated with the following primary antibodies: anti-ASS1 antibody (16210-1-AP, Proteintech), anti-COX-2 antibody (12282, CST), anti-S6 antibody (2317, CST), anti-p-S6 antibody (4858, CST), anti-STAT3 antibody (4904, CST), anti-p-STAT3 antibody (9134, CST), anti-mTOR antibody (66889- 1-Ig, Proteintech), anti-p-mTOR antibody (5536, CST), anti-ASL antibody (16645-1-AP, Proteintech), anti-IL1β antibody (66737-1-Ig, Proteintech) anti-GAPDH monoclonal antibody (HRP-60004, Proteintech), HRP-linked anti-rabbit IgG (SA00001-2, Proteintech), HRP-linked anti-mouse IgG (SA00001-1, Proteintech). .. Semiquantitative analysis of each protein was performed using ImageJ software (version 1.5), and the band densities were normalized to the band density of GAPDH.
Staining:Article Title: Monocytes Reprogrammed by 4-PBA Potently Contribute to the Resolution of Inflammation and Atherosclerosis
Article Snippet: Paraformaldehyde, Triton X-100, goat serum, rabbit anti–mouse PMP70 antibody, and Alexa Fluor 488 goat anti–rabbit secondary antibody were all supplied in the SelectFX Alexa Fluor 488 peroxisome labeling kit (Thermo Fisher Scientific, no. S34201). .. After washing with PBS, the cells were stained with CoraLite®594 conjugated anti–mTOR antibody (1:300 dilution; Proteintech, no. CL594-66888) or PE conjugated anti– SYK antibody (1:500 dilution; BioLegend, no. 646004). .. After washing with PBS, the samples were mounted with antifade mountant (Invitrogen).
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