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Image Search Results
Journal: Microbiology Spectrum
Article Title: Fatty acid synthase may facilitate the trafficking of bovine alpha herpesvirus 1 out of the Golgi apparatus, potentially promoting viral infection
doi: 10.1128/spectrum.01388-25
Figure Lengend Snippet: Detection of FASN protein in TG neurons during viral acute infection and latency by IHC. ( A ) TG tissues were collected from calves subjected to mock infection, acute infection (at 4 days post-infection), or latent infection (at 60 days post-infection). Thin sections of 10 µm were cut from formalin-fixed, paraffin-embedded TG tissues and subjected to FASN protein detection by IHC using FASN-specific polyclonal antibody (Proteintech, cat#10624-2-AP, 1:500). Biotinylated goat anti-rabbit IgG (Vector Laboratories) was used as the secondary antibody. ( B ) The percentage of FASN-positive neurons was calculated from 468 neurons of uninfected calves, 467 neurons from acutely infected calves, and 413 TG neurons from latency calves. Data shown are representative of two independent experiments. Scale bars = 100 µm.
Article Snippet:
Techniques: Infection, Formalin-fixed Paraffin-Embedded, Plasmid Preparation
Journal: Microbiology Spectrum
Article Title: Fatty acid synthase may facilitate the trafficking of bovine alpha herpesvirus 1 out of the Golgi apparatus, potentially promoting viral infection
doi: 10.1128/spectrum.01388-25
Figure Lengend Snippet: BoAHV-1 infection decreases the expression of FASN protein across various cell cultures. ( A ) MDBK cells that were confluent in 60 mm dishes were either mock infected or infected with BoAHV-1 at an MOI of 1. After 4, 8, 16, and 24 h of infection, cell lysates were prepared and analyzed by Western blotting using antibodies against FASN (Proteintech, cat# 10624-2-AP, 1:10,000) and virion-associated proteins (VMRD, cat# P170703-001, 1:5,000). ( C ) MDBK cells were either mock-infected or infected with BoAHV-1 at MOIs of 0.1, 1, and 10, respectively. At 24 hpi, cell lysates were prepared for Western blotting to assess the protein expression of FASN and virion-associated proteins. ( E ) Neuro-2A cells were either mock-infected or infected with BoAHV-1 at an MOI of 10. After 12, 24, 36, and 48 hours of infection, cell lysates were prepared and analyzed by Western blotting to detect FASN protein and virion-associated proteins. Tubulin was probed and served as a protein loading control and for subsequent quantitative analysis. ( B, D, and F ) Quantitative analysis of FASN band intensities was performed using the freeware software Image J. Changes in FASN protein levels after infection were calculated relative to mock-infected controls, which were set at 100%. ( G ) Neuro-2A cells in 24-well plates were either mock-infected or infected with BoAHV-1 at an MOI of 10. At 36 and 48 hpi, cell survival was assessed using the CCK-8 assay kit (Beyotime, Shanghai, China, cat# C0038), following the manufacturer’s instructions. ( H ) MDBK and Neuro-2A cells in six-well plates were either mock infected or infected with the virus (MOI of 1 for MDBK cells and MOI of 10 for Neuro-2A cells) for 24 h. Total RNA was then extracted from the cells for the detection of FASN mRNA using RT-qPCR. The data shown are means of three independent experiments with error bars indicating standard deviations. Significance was assessed by standard t -test (ns, not significant; * P < 0.05; ** P < 0.01; *** P < 0.001).
Article Snippet:
Techniques: Infection, Expressing, Western Blot, Control, Software, CCK-8 Assay, Virus, Quantitative RT-PCR
Journal: Microbiology Spectrum
Article Title: Fatty acid synthase may facilitate the trafficking of bovine alpha herpesvirus 1 out of the Golgi apparatus, potentially promoting viral infection
doi: 10.1128/spectrum.01388-25
Figure Lengend Snippet: Examination of FASN localization in BoAHV-1-infected MDBK cells using IFA assay. MDBK cells were seeded into 24-well plates containing coverslips and cultured until they reached 90% confluence. The cells were then either mock-infected or infected with BoAHV-1 at an MOI of 1 for 4, 16, and 24 h, respectively. The cells were immunostained with antibodies against FASN (Green; Abclonal, cat# A21182 , 1:200) and the viral protein gD (Red; VMRD, cat# 1B8-F11, 1:1,000). Nuclei were counterstained with DAPI (4′,6-diamidino-2-phenylindole; Blue). Immunofluorescence was visualized, and images were captured using confocal microscopy (Zeiss). Zoomed-in images framed in red highlight the typical co-localization of FASN with the viral protein gD. These images are representative of results from three independent experiments. Scale bars = 20 µm. ND, not done.
Article Snippet:
Techniques: Infection, Cell Culture, Immunofluorescence, Confocal Microscopy
Journal: Microbiology Spectrum
Article Title: Fatty acid synthase may facilitate the trafficking of bovine alpha herpesvirus 1 out of the Golgi apparatus, potentially promoting viral infection
doi: 10.1128/spectrum.01388-25
Figure Lengend Snippet: Examination of FASN localization in BoAHV-1-infected Neuro-2A cells using IFA assay. Neuro-2A cells were seeded into 24-well plates containing coverslips and cultured until they reached 90% confluence. The cells were then either mock-infected or infected with BoAHV-1 at an MOI of 1 for 48 and 72 h, respectively. They were immunostained with antibodies against the FASN protein (Red; Proteintech, cat# 10624-2-AP, 1:600) and the virion-associated protein (Green; VMRD, cat# P170703-001, 1:2,000). Nuclei were counterstained with DAPI (Blue). Immunofluorescence was visualized, and images were captured using confocal microscopy (Zeiss). These images are representative of results from three independent experiments. Scale bars = 20 µm.
Article Snippet:
Techniques: Infection, Cell Culture, Immunofluorescence, Confocal Microscopy
Journal: Microbiology Spectrum
Article Title: Fatty acid synthase may facilitate the trafficking of bovine alpha herpesvirus 1 out of the Golgi apparatus, potentially promoting viral infection
doi: 10.1128/spectrum.01388-25
Figure Lengend Snippet: Examination of FASN protein in the Golgi apparatus in MDBK cells. ( A ) MDBK cells were seeded into 24-well plates containing coverslips and cultured until they reached 90% confluence. The cells were then either mock-infected or infected with BoAHV-1 at an MOI of 1 for 24 h. They were immunostained with antibodies against the FASN protein (Green; Proteintech, cat# 10624-2-AP, 1:600) and GP73 (Red; Proteintech, cat# 66331-1-1g, 1:200). Nuclei were counterstained with DAPI (Blue). Immunofluorescence was visualized, and images were captured using confocal microscopy (Zeiss). These images are representative of results from three independent experiments. Scale bars = 20 µm. ( B ) MDBK cells in 100 mm dishes were either mock-infected or infected with BoAHV-1 (MOI = 1) for 24 h. The cells were then collected to isolate the Golgi fraction using a commercial kit (Beijing Biolabo Technology, cat# HR0247-50T), following the manufacturer’s protocol. The lysates were subjected to Western blot analysis using antibodies against FASN (Proteintech, cat# 10624-2-AP, 1:10,000) and GOLGA1 (Abclonal, cat# A14688, 1:1,000). GOLGA1 served as a marker for the Golgi apparatus and as a protein loading control. The data presented are representative of three independent experiments. ( C ) Quantitative analysis of FASN band intensities was performed using the freeware software Image J. Changes in FASN protein levels after infection were calculated relative to mock-infected controls, which were set at 100%. The data shown are means of three independent experiments with error bars indicating standard deviations. Significance was assessed by a standard t -test (** P < 0.01).
Article Snippet:
Techniques: Cell Culture, Infection, Immunofluorescence, Confocal Microscopy, Western Blot, Marker, Control, Software
Journal: Microbiology Spectrum
Article Title: Fatty acid synthase may facilitate the trafficking of bovine alpha herpesvirus 1 out of the Golgi apparatus, potentially promoting viral infection
doi: 10.1128/spectrum.01388-25
Figure Lengend Snippet: Determine the roles of FASN in BoAHV-1 productive infection in MDBK cells using transfection of siRNAs. ( A ) MDBK cells in six-well plates were transfected with either scrambled siRNA (150 pmol) or individual siRNAs targeting FASN (150 pmol), referred to as siRNA1, siRNA2, siRNA3, and siRNA4, respectively. At 48 h after transfection, cell lysates were prepared and subjected to Western blot analysis to detect FASN protein levels using an antibody against FASN (Proteintech, cat# 10624-2-AP, 1:10,000). ( C and E ) MDBK cells in six-well plates were transfected with either scrambled siRNA (150 pmol) or the indicated siRNAs (150 pmol). At 36 hpi, the cells were infected with BoAHV-1 at an MOI of 1. After 24 h of infection, the cells were collected and subjected to Western blot analysis using antibodies against FASN (Proteintech, cat# 10624-2-AP, 1:10,000) ( C ) and virus-associated proteins (VMRD, cat# P170703-001, 1:5,000) ( E ). ( B, D, and F ) The band intensities were quantified using the free software Image J. The intensity of each band was first normalized to that of the respective loading control Tubulin, and then normalized to that of the control transfected with scramble siRNA, which was arbitrarily set to 100%. ( G ) MDBK cells in six-well plates were transfected with either scrambled siRNA (150 pmol) or the indicated siRNAs (150 pmol). At 36 h post-transfection, the cells were infected with the virus at an MOI of 1 for 24 h. Subsequently, total RNA was extracted from the cells for the detection of viral mRNA using RT-qPCR. Primers specific for bICP4, bICP27, viral DNA polymerase, and viral protein gC were used, respectively. The data shown are means of three independent experiments, with error bars representing standard deviations. Significance was assessed using a Student’s t -test (ns, not significant; * P < 0.05; ** P < 0.01; *** P < 0.001).
Article Snippet:
Techniques: Infection, Transfection, Western Blot, Virus, Software, Control, Quantitative RT-PCR
Journal: Microbiology Spectrum
Article Title: Fatty acid synthase may facilitate the trafficking of bovine alpha herpesvirus 1 out of the Golgi apparatus, potentially promoting viral infection
doi: 10.1128/spectrum.01388-25
Figure Lengend Snippet: Determine the roles of FASN in BoAHV-1 productive infection in MDBK cells using a chemical inhibitor. ( A and C ) MDBK cells in six-well plates were infected with BoAHV-1 (MOI = 1) and treated either with DMSO control or with the FASN-specific inhibitor Cerulenin at the indicated concentrations. At 24 hpi, the cells were collected either to prepare cell lysates for Western blotting using an antibody against virus-associated proteins (VMRD, cat# P170703-001, 1:5,000) ( A ) or to extract DNA for subsequent qPCR analysis using gB-specific primers ( C ). ( B ) The intensity of bands for virus-associated proteins was quantified using the freeware software Image J. Changes in the levels of individual bands upon treatment with the inhibitor were calculated relative to those of the DMSO control, which was set at 100%. ( D ) Virus titers in the supernatants were determined and expressed as TCID 50 /mL. ( E ) MDBK cells in six-well plates were infected with the virus at an MOI of 1 for 24 h and treated with either DMSO control or 10 µM Cerulenin. Total RNA was then extracted from the cells for the detection of viral mRNA using RT-qPCR. Primers specific for bICP27, viral DNA polymerase, and viral protein gC were used, respectively. The data shown are means of three independent experiments, with error bars representing standard deviations. Significance was assessed by a standard t -test (ns, not significant; * P < 0.05; ** P < 0.01; *** P < 0.001).
Article Snippet:
Techniques: Infection, Control, Western Blot, Virus, Software, Quantitative RT-PCR
Journal: Microbiology Spectrum
Article Title: Fatty acid synthase may facilitate the trafficking of bovine alpha herpesvirus 1 out of the Golgi apparatus, potentially promoting viral infection
doi: 10.1128/spectrum.01388-25
Figure Lengend Snippet: Determine the roles of FASN in BoAHV-1 productive infection in Neuro-2A cells using both siRNA and chemical inhibitor. ( A ) Neuro-2A cells in six-well plates were transfected with either scrambled siRNA (150 pmol) or individual siRNAs targeting FASN (150 pmol), referred to as siRNA1, siRNA2, siRNA3, and siRNA4, respectively. At 48 h post-transfection, cell lysates were prepared and subjected to Western blot analysis to detect FASN protein levels using an antibody against FASN (Proteintech, cat# 10624-2-AP, 1:10,000). ( C ) Neuro-2A cells in six-well plates were transfected with either scrambled siRNA (150 pmol) or siRNA1 (150 pmol). At 36 h post-transfection, the cells were infected with BoAHV-1 at an MOI of 10. After 24 h of infection, the cells were collected and subjected to Western blot analysis using a monoclonal antibody against viral protein gC (VMRD, cat#F2, 1:2,000). ( E ) Neuro-2A cells in six-well plates were infected with BoAHV-1 (MOI = 10) and treated either with DMSO control or with the FASN-specific inhibitor Cerulenin at the indicated concentrations. At 24 hpi, the cell lysates were prepared and subjected to Western blotting to detect viral protein gC. ( B, D, and F ) The band intensities were quantified using the free software Image J. The intensity of each band was first normalized to that of the respective loading control Tubulin, and then normalized to that of either DMSO-treated or scrambled siRNA-transfected control, which was arbitrarily set to 100%. The data shown are means of three independent experiments, with error bars representing standard deviations. Significance was assessed using a Student’s t -test (ns, not significant; * P < 0.05; ** P < 0.01).
Article Snippet:
Techniques: Infection, Transfection, Western Blot, Control, Software
Journal: Microbiology Spectrum
Article Title: Fatty acid synthase may facilitate the trafficking of bovine alpha herpesvirus 1 out of the Golgi apparatus, potentially promoting viral infection
doi: 10.1128/spectrum.01388-25
Figure Lengend Snippet: Determine the effects of FASN inhibitor, Cerulenin, on the accumulation of gD in the Golgi apparatus. ( A ) Diagram shows the treatment manner of virus-infected cells by both Cerulenin and Monensin. ( B ) MDBK cells of either mock-infected or virus-infected were treated either with vehicle control DMSO or with Cerulenin (10 µM) or Monensin (10 µM) for a duration of 4 h prior to the termination of infection. At 24 hpi, the cells were collected to purify the Golgi apparatus using a commercial purification kit (Beijing Biolabo Technology, cat# HR0247-50T). Subsequently, Western blot analysis was performed to detect the protein levels of gD in the Golgi fractions. GOLGA1, a marker of the Golgi apparatus, was used as a loading control and for subsequent quantitative analysis. ( C ) The band intensity was analyzed with the free software Image J. The intensity of each band was first normalized to that of the respective loading control GOLGA1, and then normalized to that of the control treated with DMSO, which was arbitrarily set to 1. The data shown are means of three independent experiments with error bars indicating standard deviations. Significance was assessed with a Student’s t -test (* P < 0.05, ** P < 0.01).
Article Snippet:
Techniques: Virus, Infection, Control, Purification, Western Blot, Marker, Software
Journal: International Journal of Molecular Sciences
Article Title: Molecular Insights into the Inhibition of Lipid Accumulation in Hepatocytes by Unique Extracts of Ashwagandha
doi: 10.3390/ijms252212256
Figure Lengend Snippet: Effects of withanolides and Ashwagandha extracts on key regulators of lipid metabolism. ( A ) Western blot showing the effect of Withaferin A (Wi-A) and Withanone (Wi-N) on fatty acid synthase (FASN) protein level in Huh7 cells treated with PA. Quantitation from three independent experiments is shown below. ( B ) Western blot displaying the effect of Wi-A and Wi-N on peroxisome proliferator-activated receptor (PPAR)γ protein levels in palmitic acid (PA)-treated Huh7 cells. Quantitation from three independent experiments is shown below. ( C ) Western blot analysis showing the effect of Ashwagandha extracts on PPARγ protein levels in Huh7 cells treated with PA. Quantitation from three independent experiments is shown below. For the quantitation graphs, data were normalized against the PA group and plotted as fold differences (mean ± SD; n = 3). The statistical significance of the data (mean ± SD; n = 3; and * p < 0.05, ** p < 0.01, and *** p < 0.001) was calculated using an unpaired Student’s t -test.
Article Snippet: The membranes were blocked with 3% bovine serum albumin (BSA) at room temperature for 1 h. The blocked membranes were probed with the following target-protein-specific primary antibodies at 4 °C overnight: CARF (raised in our lab [ ]), SREBP-1c (sc-13551, Santa Cruz Biotechnology, Paso Robles, CA, USA),
Techniques: Western Blot, Quantitation Assay
Journal: International Journal of Molecular Sciences
Article Title: Molecular Insights into the Inhibition of Lipid Accumulation in Hepatocytes by Unique Extracts of Ashwagandha
doi: 10.3390/ijms252212256
Figure Lengend Snippet: Effects of Ashwagandha extracts on the de novo lipogenesis pathway. ( A ) A schematic representation of the de novo lipogenesis pathway, key enzymes, and steps involved in fatty acid synthesis is shown. ( B ) Western blot showing the effects of Ashwagandha extracts on precursor and mature sterol regulatory element-binding protein-1c (SREBP-1c) expression levels in Huh7 cells treated with PA. β-actin was used as a loading control. Quantitation from three independent experiments is shown below. ( C ) Ashwagandha extracts decreased SREBP-1c mRNA and its target genes Acetyl-CoA Carboxylase 1 ( ACC1 ), fatty acid synthase ( FASN ) and stearoyl-CoA desaturase 1 ( SCD1 ) in PA-treated HepG2 cells. Data were normalized against the PA group for all panels and plotted as fold differences (mean ± SD; n = 3). The statistical significance of the data (mean ± SD; n = 3; and * p < 0.05, ** p < 0.01, and *** p < 0.001) was calculated using an unpaired Student’s t -test.
Article Snippet: The membranes were blocked with 3% bovine serum albumin (BSA) at room temperature for 1 h. The blocked membranes were probed with the following target-protein-specific primary antibodies at 4 °C overnight: CARF (raised in our lab [ ]), SREBP-1c (sc-13551, Santa Cruz Biotechnology, Paso Robles, CA, USA),
Techniques: Western Blot, Binding Assay, Expressing, Control, Quantitation Assay
Journal: Nutrients
Article Title: Improvement of Hypertriglyceridemia by Roasted Nelumbinis folium in High Fat/High Cholesterol Diet Rat Model
doi: 10.3390/nu12123859
Figure Lengend Snippet: Hematological changes in the treatment of roasted Nelumbo nucifera (RN) in high fat/cholesterol (HFC) diet Rats. Representative plasma images of HFC diet-fed rats appear turbid compared to those of the control group ( A ). Representative plasma image of HFC diet rats that appear turbid compared to the control group ( A ). Measurements of plasma insulin and glucose ( B ) TG, T-Cho, LDL, and HDL ( C ); FFA and FAS ( D ). CON, regular diet; HFC, HFC diet; FLU, fluvastatin (3 mg/kg/day) with HFC diet; RNN, RN (100 mg/kg/day) with HFC diet; RNH, RN (200 mg/kg/day) with HFC diet; TG, triglyceride; T-Cho, total cholesterol; LDL, low-density lipoprotein; HDL, high-density lipoprotein; FFA, free fatty acid; FAS, fatty acid synthase. The values represent the means ± standard error. ** p <0.01 and *** p < 0.001 vs. CON; # p < 0.05, ## p < 0.01, and ### p < 0.001 vs. HFC.
Article Snippet: The levels of fatty acids (ab65341, Abcam Inc., Bristol, UK),
Techniques: Clinical Proteomics, Control
Journal: The Journal of Biological Chemistry
Article Title: Fatty acid synthase inhibits the O- GlcNAcase during oxidative stress
doi: 10.1074/jbc.M116.760785
Figure Lengend Snippet: Oxidative stress induces the association of OGA with FAS, FLNA, HSC70, and OGT. U2OS cells were treated with Vehicle (V) or H2O2 (2.5 mm, 1–3 h). n = 3. A, anti-OGA antibody (IP: OGA; top panel) or a rabbit isotype control immunoglobulin (IP: IgG; middle panel) was used to enrich endogenous OGA from NETN cell lysates (500 μg), of which 1.5–2% (input) and 30–40% (immunoprecipitate) were analyzed by SDS-PAGE. OGA, FAS, FLNA, HSC70, OGT (positive control), and actin (loading/negative control) were detected by Western blotting. B, anti-FAS antibody (IP: FAS; top panel) or a rabbit isotype control immunoglobulin (IP: IgG; middle panel) was used to enrich endogenous FAS from NETN cell lysates (250 μg), of which 3% (input) and 60% (immunoprecipitate) were analyzed by SDS-PAGE. FAS, OGA, HSC70, OGT, and actin (loading/negative control) were detected by Western blotting. C, U2OS cells were transfected with pcDNA3.1 (control) or pCMV-SPORT6 V5-FAS (test). An anti-V5 antibody was used to enrich V5-FAS from control and test NETN cell lysates (300 μg), of which 1.7% (input) and 33.3% (immunoprecipitate) were analyzed by SDS-PAGE. V5, OGA, HSC70, OGT, and actin (loading/negative control) were detected by Western blotting. A–C, FAS (CST) and FAS (NB) represent anti-FAS antibody from Cell Signaling Technology and Novus Biologicals, respectively. To ensure that images were in the linear range, Western blot exposures from the input and immunoprecipitated fractions are often different. The exposure lengths for the test and control isotype antibody immunoprecipitates are always identical. The migration of molecular mass (MW) markers is indicated.
Article Snippet: The following antibodies were used for Western blotting analysis: anti-OGA (345), anti-O-GlcNAc (CTD110.6) (gift from The Johns Hopkins University School of Medicine Core C4); anti-FLNA (A301-135A; Bethyl Laboratories); anti-HA (H6908), anti-actin (A5060), anti-OGT (DM17, O6264), anti-mouse IgM-HRP (A8786; Sigma); NeutrAvidin-HRP (31030), anti-V5 (461157; Thermo Fisher Scientific); anti-Myc (9E10, CRL-1729; American Type Culture Collection (ATCC), Manassas, VA); anti-FAS (3180; Cell Signaling Technology, Danvers, MA);
Techniques: SDS Page, Positive Control, Negative Control, Western Blot, Transfection, Immunoprecipitation, Migration