srebp1 antibody - bsa free  (Bio-Techne corporation)

Journal: Animal Nutrition

Article Title: Influences of dietary guanidinoacetic acid supplementation on performance and proteins involved in milk fat and protein synthesis in dairy cows

doi: 10.1016/j.aninu.2025.07.008

Figure Lengend Snippet: Changes in the expression levels of proteins concerned with fatty acid synthesis. (A) Western blot analysis of AMPK, p-AMPK, PPARγ, SREBP1, ACACA, p-ACACA, FASN, and SCD1 in bovine mammary gland tissues treated with 0 g/kg (control) and 0.36 g/kg DM of GAA derived from RPGAA (MRPGAA); β-actin was used as the loading control. (B) Mean ± SEM of immunopositive bands of PPARγ, SREBP1, p-ACACA/ACACA, FASN, SCD1, and p-AMPK/AMPK. AMPK = adenosine monophosphate activated protein kinase; PPARγ = peroxisome proliferator-activated receptor gamma; SREBP1 = sterol regulatory element-binding protein 1; ACACA = acetyl-coenzyme A carboxylase-α; FASN = fatty acid synthase; SCD1 = stearoyl-CoA desaturase 1; GAA = guanidinoacetic acid; RPGAA = rumen-protected guanidinoacetic acid; p = phosphorylated. ∗ P < 0.05 and ∗∗ P < 0.01 versus the control group.

Article Snippet: Rabbit anti-sterol regulatory element-binding protein 1 (SREBP1; #NB100-2215) was purchased from Novus Biologicals (Centennial, CO, USA).

Techniques: Expressing, Western Blot, Control, Derivative Assay, Binding Assay

Journal: Microbiome

Article Title: Altered rumen bacterial flora is associated with increased lipogenesis of adipose tissue in obese dairy cows before calving

doi: 10.1186/s40168-026-02343-7

Figure Lengend Snippet: Differential lipid and glucose metabolism of adipose tissue and blood FFA profiles between normal and obese dairy cows. A Representative images of hematoxylin–eosin (H&E) staining of the adipose tissue. B , C Adipocyte diameter and TG content in the adipose tissue. D Representative blots of SREBP-1C, p-HSL, HSL, ATGL, p-AKT, AKT, and ACTB. E Quantification of protein levels of SREBP-1C, p-HSL, ATGL, and p-AKT. F Heatmap of the abundance of genes related to lipid and glucose metabolism in the adipose tissue. G The PCoA analysis based on plasma FFA profiles. H Average concentrations of plasma FFA. I Heatmap of the relative concentrations of plasma FFA profiles. J Significantly different FFA in plasma. n = 10 per group. Significant differences were tested using the independent samples t test. Data with error bars were expressed as mean ± SD. * P < 0.05, ** P < 0.01

Article Snippet: After being blocked with 3% BSA for 4 h at room temperature, membranes were incubated overnight at 4 °C with specific antibodies against protein sterol regulatory element-binding protein 1C (SREBP-1C; 1:1,000; NB100–2215; Novus Biologicals, Littleton, CO, USA), hormone-sensitive lipase (HSL; 1:1,000; AF6403; Affinity Biosciences Ltd., Jiangsu, China), phosphorylated-HSL (p-HSL; 1:1,000; AF2350; Affinity Biosciences Ltd.), adipose triglyceride lipase (ATGL; 1:1,000; Ab99532; Abcam, Cambridge, MA, USA), protein kinase B (AKT; 1:1,000; cat. no. 9272; Cell Signaling Technology Inc., MA, USA), p-AKT (1:1,000; cat. no. 4060; Cell Signaling Technology Inc.), and ACTB (1:2,000; Ab8226; Abcam), respectively.

Techniques: Staining, Clinical Proteomics

Journal: Microbiome

Article Title: Altered rumen bacterial flora is associated with increased lipogenesis of adipose tissue in obese dairy cows before calving

doi: 10.1186/s40168-026-02343-7

Figure Lengend Snippet: A schematic illustration that depicts the association between rumen bacterial flora, functions and metabolites, and adipocyte lipid deposition in obese dairy cows. The rumen of prepartum obese dairy cows exhibited a higher abundance of VFA-producing bacteria and an increased capacity for carbohydrate degradation and VFA formation. The acetate and propionate produced in the rumen were utilized by the liver for glucose and TG synthesis. Moreover, the elevated levels of circulating glucose and FFA contributed to lipid accumulation in adipocytes, leading to obesity in dairy cows. VFA volatile fatty acids, FFA free fatty acids, TG triglyceride, LD lipid droplet, SREBP - 1C sterol regulatory element-binding protein 1C, ACSS2 acyl-coA synthetase short chain family member 2, CD36 cluster of differentiation 36, PCK1 phosphoenolpyruvate carboxykinase 1, PC pyruvate carboxylase, G6P1 glucose-6-phosphatase 1, GLU4 glucose transporter type, PLIN1 perilipin 1, CIDEC cell death-inducing DNA fragmentation factor-α-like effector c, p - HSL phosphorylated-hormone-sensitive lipase, ATGL adipose triglyceride lipase. Red arrow and font indicate upregulation in prepartum obese cows

Article Snippet: After being blocked with 3% BSA for 4 h at room temperature, membranes were incubated overnight at 4 °C with specific antibodies against protein sterol regulatory element-binding protein 1C (SREBP-1C; 1:1,000; NB100–2215; Novus Biologicals, Littleton, CO, USA), hormone-sensitive lipase (HSL; 1:1,000; AF6403; Affinity Biosciences Ltd., Jiangsu, China), phosphorylated-HSL (p-HSL; 1:1,000; AF2350; Affinity Biosciences Ltd.), adipose triglyceride lipase (ATGL; 1:1,000; Ab99532; Abcam, Cambridge, MA, USA), protein kinase B (AKT; 1:1,000; cat. no. 9272; Cell Signaling Technology Inc., MA, USA), p-AKT (1:1,000; cat. no. 4060; Cell Signaling Technology Inc.), and ACTB (1:2,000; Ab8226; Abcam), respectively.

Techniques: Bacteria, Produced, Binding Assay

Journal: Advanced Science

Article Title: Inhibition of Tumor Lipogenesis and Growth by Peptide‐Based Targeting of SREBP Activation

doi: 10.1002/advs.202508111

Figure Lengend Snippet: The Insig1/2 loop 1 peptide binds to S90‐phosphorylated PCK1 and blocks PCK1‐mediated SREBP activation and tumor cell proliferation. Immunoblotting analyses were performed using the indicated antibodies; representative results from three independent experiments are shown (IP, immunoprecipitation; WB, western blot). a) Huh7 cells expressing various Insig1 (top) or Insig2 (bottom) truncation mutants were treated with or without IGF1 (100 ng mL −1 ) for 1 h. b,c) Biolayer interferometry assays (Sartorius Octet) were employed to measure the binding affinity of Insig1/2 loop 1 peptide to GST‐PCK1 or GST‐PCK1 pS90 proteins. Data represent three independent experiments. d) Flag‐PCK1‐expressing Huh7 cells were pretreated with Insig1/2 loop 1 peptide (15 µ m ) for 1 h before IGF1 (100 ng mL −1 ) stimulation. e) Huh7 cells were pretreated with low (7.5 µ m ) or high (15 µ m ) concentrations of Insig1/2 loop 1 peptide for 1 h before IGF1 (100 ng mL −1 ) stimulation. f) Huh7 cells transiently transfected with β‐galactosidase and SRE‐driven luciferase reporter vectors were pretreated with Insig1/2 loop 1 peptide (15 µM) for 1 h, followed by stimulation with or without IGF1 (100 ng mL −1 ) for 14 h. SRE luciferase activity was normalized to β‐galactosidase activity. Data represent the mean ± SD (n = 3). Statistical significance was determined by one‐way analysis of variance (ANOVA). g) Following pretreatment with Insig1/2 loop 1 peptide (15 µ m ) and IGF1 stimulation (100 ng mL −1 , 8 h), Huh7 cells were subjected to immunofluorescence analysis (scale bar: 10 µm). h) Quantitative PCR analysis of SREBP1 target gene GPAM in Huh7 cells pretreated with Insig1/2 loop 1 peptide (15 µ m ) and stimulated with IGF1 (100 ng mL −1 ) for 10 h. Data represent the mean ± SD (n = 3). Statistical significance was determined by one‐way analysis of variance (ANOVA). i) Quantitative PCR analysis of SREBP1 target gene FASN in Huh7 cells pretreated with Insig1/2 loop 1 peptide (15 µ m ) and stimulated with IGF1 (100 ng mL −1 ) for 10 h. Data represent the mean ± SD (n = 3). Statistical significance was determined by one‐way analysis of variance (ANOVA). j) Intracellular triglyceride (TG) content was quantified in Huh7 cells pretreated with Insig1/2 loop 1 peptide (25 µ m ) and stimulated with IGF1 (100 ng mL −1 ) for 24 h. Data represent the mean ± SD (n = 3). Statistical significance was determined by one‐way analysis of variance (ANOVA). k) Cell proliferation was assessed by automated cell counting after 72 h of IGF1 stimulation (100 ng mL −1 ) in Huh7 cells pretreated with Insig1/2 loop 1 peptide (25 µ m ) for 1 h. Data represent the mean ± SD (n = 3). Statistical significance was determined by one‐way analysis of variance (ANOVA). l) FLAG‐Insig1, Flag‐Insig1 S207A or FLAG‐ Insig1 S207D was expressed in Huh7 cells with depletion of Insig1 by expressing Insig1 shRNA. Quantitative PCR analysis of SREBP1 target gene GPAM in Huh7 cells pretreated with Insig1/2 loop 1 peptide (15 µ m ) and stimulated with IGF1 (100 ng mL −1 ) for 10 h. Data represent the mean ± SD (n = 3). Statistical significance was determined by two‐way analysis of variance (ANOVA). m) FLAG‐Insig1, Flag‐Insig1 S207A or FLAG‐ Insig1 S207D was expressed in Huh7 cells with depletion of Insig1 by expressing Insig1 shRNA. Quantitative PCR analysis of SREBP1 target gene FASN in Huh7 cells pretreated with Insig1/2 loop 1 peptide (15 µ m ) and stimulated with IGF1 (100 ng mL −1 ) for 10 h. Data represent the mean ± SD (n = 3). Statistical significance was determined by two‐way analysis of variance (ANOVA). * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; ns, no significance.

Article Snippet: For immunofluorescence and IHC analyses, SREBP1 (NB100‐2215) and SREBP2 (NB100‐74543) antibodies were sourced from Novus.

Techniques: Activation Assay, Western Blot, Immunoprecipitation, Expressing, Binding Assay, Transfection, Luciferase, Activity Assay, Immunofluorescence, Real-time Polymerase Chain Reaction, Cell Counting, shRNA

Journal: Advanced Science

Article Title: Inhibition of Tumor Lipogenesis and Growth by Peptide‐Based Targeting of SREBP Activation

doi: 10.1002/advs.202508111

Figure Lengend Snippet: Insig1/2 loop 1 peptide treatment inhibits SREBP1 activity and lipid synthesis and induces tumor cell apoptosis in liver tumors. a) Immunohistochemical (IHC) analysis of mouse tumor specimens were performed using the indicated antibodies. Representative micrographs are shown (scale bar: 20 µm). b,c) Cryosections of mouse tumor tissues were prepared for histological analysis. (b) Lipid accumulation was assessed by Oil Red O staining. Representative micrographs are shown (scale bar: 20 µm). (c) Apoptotic cells were detected by TUNEL assay. Representative micrographs are shown (scale bar: 20 µm). d,e) Quantitative analysis of (d) apoptotic areas (TUNEL‐positive regions) and (e) lipid droplet counts (Oil Red O‐positive particles) was performed. Data represent the mean ± SD (n = 4). Statistical significance was determined by one‐way analysis of variance (ANOVA). ** p < 0.01, **** p < 0.0001, ns, no significance.

Article Snippet: For immunofluorescence and IHC analyses, SREBP1 (NB100‐2215) and SREBP2 (NB100‐74543) antibodies were sourced from Novus.

Techniques: Activity Assay, Immunohistochemical staining, Staining, TUNEL Assay

Journal: Advanced Science

Article Title: Inhibition of Tumor Lipogenesis and Growth by Peptide‐Based Targeting of SREBP Activation

doi: 10.1002/advs.202508111

Figure Lengend Snippet: Mechanistic insights into tumor growth inhibition by Insig1/2 loop1 peptide through PCK1‐mediated SREBP1 activation and lipogenesis regulation. Schematic diagram of the peptide‐mediated tumor proliferation inhibition mechanism. Created in https://BioRender.com .

Article Snippet: For immunofluorescence and IHC analyses, SREBP1 (NB100‐2215) and SREBP2 (NB100‐74543) antibodies were sourced from Novus.

Techniques: Inhibition, Activation Assay