anti creb ab  (OriGene)

Journal: Journal of Biomedical Research

Article Title: RAF1 in AgRP neurons involved in the regulation of energy metabolism via the MAPK signaling pathway

doi: 10.7555/JBR.39.20250114

Figure Lengend Snippet: Raf1 regulated MAPK signaling under insulin stimulation. A: Relative mRNA levels of Raf1 , Agrp , and Npy in the hypothalamus of control and AgRP- Raf1 -OE mice fed an NCD ( n = 5–6 mice). B: Western blotting analysis of protein levels of FLAG, MEK1/2, pMEK1/2, ERK1/2, pERK1/2, CREB, and pCREB in the N42 Raf1 -overexpression cells. β-Actin served as the internal control ( n = 3). C: Phosphorylation levels of MEK1/2, ERK1/2, and CREB proteins in the N42 Raf1 -overexpression cells. D: Western blotting analysis of protein levels of RAF1, MEK1/2, pMEK1/2, ERK1/2, pERK1/2, and pCREB in the N42 Raf1 -knockout cells. β-Actin served as the internal control ( n = 3). E: Phosphorylation levels of MEK1/2, ERK1/2, and CREB proteins in the N42 Raf1 -knockout cells. F: Representative IF staining of pCREB in AgRP neurons of control and AgRP- Raf1 -OE mice following 2 mU insulin (icv) stimulation ( n = 3 mice; scale bars, 100 μm). G: Fluorescence intensity quantification of pCREB co-localized with HA/mCherry ( n = 3 mice; AAV-DIO-mCherry, N = 77; AAV-DIO- Raf1 -HA, N = 105). N represents the cell number, and n represents the mouse number. Data are presented as the mean ± standard error of the mean. * P < 0.05, ** P < 0.01, and *** P < 0.001 by unpaired t -tests (A, C, E, and G). Abbreviations: AgRP, agouti-related peptide; CREB, cAMP response element-binding protein; ERK1/2, extracellular signal-regulated kinases 1 and 2; icv, intra-cerebroventricular injection; IF, immunofluorescence; MEK1/2, mitogen-activated protein kinase kinases 1 and 2; NCD, normal chow diet; NPY, neuropeptide Y; OE, overexpression; p-CREB, phospho-CREB; p-ERK1/2, phospho-ERK1/2; p-MEK1/2, phospho-MEK1/2; RAF1, v-raf-leukemia viral oncogene 1.

Article Snippet: The membranes were blocked with 5% non-fat milk and incubated overnight at 4 °C with primary antibodies, including mouse anti-RAF1 mAb (1∶1000, Cat. #66592-1-Ig, Proteintech, Wuhan, China), rabbit anti-ERK1/2 polyclonal antibody (1∶500, Cat. #BS2265, Bioworld, China), rabbit anti-ERK1/2 (phospho-T202/Y204) polyclonal antibody (1∶5000, Cat. #AP0484, Bioworld), rabbit anti-MEK1/2 polyclonal antibody (1∶500, Cat. #BS3599, Bioworld), rabbit anti-MEK1/2 (phospho-S2218/222) polyclonal antibody (1∶500, Cat. #BS4733, Bioworld), rabbit anti-CREB1 polyclonal antibody (1∶1000, Cat. #12208-1-AP, Proteintech), anti-CREB (phospho S133) (1∶1000, Cat. #ab32096, Abcam, Cambridge, UK), and mouse anti-β-actin mAb (1∶5000, Cat. #BS6007M, Bioworld).

Techniques: Control, Western Blot, Over Expression, Phospho-proteomics, Knock-Out, Staining, Fluorescence, Binding Assay, Injection, Immunofluorescence

Journal: iScience

Article Title: The sperm quality in DIO male mice is linked to the NF-κB signaling and Ppp2ca expression in the hypothalamus

doi: 10.1016/j.isci.2025.112110

Figure Lengend Snippet: RNA-seq coupled with bioinformatic analysis of ARC enriched with KNDy neurons in DIO male mice (A) Schematic illustration showed laser capture microdissection (LCM) of ARC enriched of Kiss1 neurons. (B) Representative fluorescence image showed the area of ARC enriched of Kiss1 neurons and bright-field image of ARC after LCM, scale bar: 100 μm. (C) Principal-component analysis (PCA) analysis of the RNA-seq data from both NCD group and HFD group. (D) Heatmap of differentially expressed genes (DEGs). (E) Distribution of DEGs in the NCD and HFD groups. (F) Volcano map of DEGs. (G) The top 20 enrichment circles of all gene ontology (GO) terms. (H) The pathway-DEGs network, was constructed based on signal transduction pathway and endocrine system pathway. (I) The distribution of 42 candidate DEGs patway sets including endocrine system, nervous system, and signal tranduction. (J) The protein interaction network of 42 candidate DEGs is represented by circles, with larger circles indicating more protein interactions. Red: upregulated genes, green: downregulated genes, gray: non-significant difference expressed genes. (K–M) The downregulated gene set was regulated by cAMP responsive element binding protein 1 (CREB1) (K), Akt kinase (AKT) (L), and RELA (M) in HFD male mice. (N) The upregulated gene set was regulated by inflammatory activation in HFD male mice.

Article Snippet: CREB1 Rabbit Polyclonal Antibody , Proteintech , Cat# 12208-1-AP; RRID: AB_2245417.

Techniques: RNA Sequencing, Laser Capture Microdissection, Fluorescence, Construct, Transduction, Binding Assay, Activation Assay

Journal: iScience

Article Title: The sperm quality in DIO male mice is linked to the NF-κB signaling and Ppp2ca expression in the hypothalamus

doi: 10.1016/j.isci.2025.112110

Figure Lengend Snippet: The elevated expression of PP2Ac and decreased activities of AKT and CREB1 in the hypothalamus were confirmed in both DIO mice and ARC IKKβ CA mice (A) The representative fluorescence images shows the expression of PP2Ac (Red) in ARC of Kiss1-cre::R26R-EYFP mice under NCD and HFD. The white arrow indicates the expression of PP2Ac in Kiss1 neurons, scale bar: 20 μm. (B) The counting statistics of PP2Ac-positive cells were performed for Kiss1 neurons in the NCD and HFD group ( n = 3). (C) Relative quantification of PP2Ac in Kiss1 neurons in the NCD and HFD group ( n = 3). (D) PP2Ac expression in ARC of ARC Control and ARC Ikkβ CA group male mice, The white arrow indicates the expression of PP2Ac in AAV-infected neurons, scale bar: 20 μm. (E and F) Western blot analysis of the protein levels of PP2Ac, AKT, pAKT, CREB1, and pCREB1 in the hypothalamus of NCD mice and HFD mice (E) and ARC Control and ARC Ikkβ CA mice (F). (G–I) Quantification of protein levels of PP2Ac (G), pAKT (H), and pCREB1 (I) in the hypothalamus of NCD and HFD mice and ARC Control and ARC Ikkβ CA mice ( n = 6). Data are presented as mean ± SEM,∗ indicates a significant difference (∗ means p < 0.05, ∗∗ means p < 0.01, ∗∗∗ means p < 0.001), “ns” indicates non-significant difference, Student’s t test.

Article Snippet: CREB1 Rabbit Polyclonal Antibody , Proteintech , Cat# 12208-1-AP; RRID: AB_2245417.

Techniques: Expressing, Fluorescence, Quantitative Proteomics, Control, Infection, Western Blot

Journal: iScience

Article Title: The sperm quality in DIO male mice is linked to the NF-κB signaling and Ppp2ca expression in the hypothalamus

doi: 10.1016/j.isci.2025.112110

Figure Lengend Snippet: The activation of NF-κB signaling or the overexpression of Ppp2ca led to the inhibition of AKT and CREB1 activities (A) Western blot analysis of Ikkβ - HA, PP2Ac, P65, and pP65 in control and Ikkβ CA cells. (B–E) The quantification of protein levels of pP65 (B), PP2Ac (C), pAKT (D), and pCREB1 (E) in control and Ikkβ CA cells ( n = 3). (F and M) Relative mRNA levels of Fos , Fosb , Fosl2 , Egr1 , and Kiss1 in control and Ikkβ CA cells (F), control and Ppp2ca -OE (M) treated cells ( n = 3). (G) Western blot analysis of AKT, pAKT, CREB1, and pCREB1 in control and Ikkβ CA cells, control and Ppp2ca -OE cells and DMSO treated and Artemisinin treated cells. (H–J, and L) The quantification of protein levels of pAKT, pCREB1 in control and Ppp2ca -OE cells (H and I) and DMSO treated and Artemisinin treated cells (J and L) ( n = 3). (K) Western blot analysis of AKT, pAKT, CREB1, and pCREB1 in the Ikkβ CA cells treated with NC, si Ppp2ca , DMSO, and SC79, as well as in Ppp2ca- OE cells treated with DSMO and SC79. (N–P) The quantification of protein levels of pAKT, and pCREB1 in the Ikkβ CA cells , treated with NC and si Ppp2ca (N) ( n = 3), DMSO and SC79 (O) ( n = 3), and in the Ppp2ca -OE cells treated with DSMO and SC79 (P) ( n = 3). Data are presented as mean ± SEM,∗ indicates a significant difference (∗ means p < 0.05, ∗∗ means p < 0.01, ∗∗∗ means p < 0.001, ∗∗∗∗ means p < 0.0001), Student’s t test.

Article Snippet: CREB1 Rabbit Polyclonal Antibody , Proteintech , Cat# 12208-1-AP; RRID: AB_2245417.

Techniques: Activation Assay, Over Expression, Inhibition, Western Blot, Control

Journal: iScience

Article Title: The sperm quality in DIO male mice is linked to the NF-κB signaling and Ppp2ca expression in the hypothalamus

doi: 10.1016/j.isci.2025.112110

Figure Lengend Snippet: Excessive expression of Ppp2ca in the ARC resulted in disturbances to LH pulse patterns and a decline in sperm quality (A) Schematic depiction of stereotaxic injection of AAV-CAG-EGFP and AAV-CAG-EGFP- Ppp2ca viral vectors into the mouse hypothalamus. (B) The ventral view of AAV virus injected mouse brains under fluorescence microscope, scale bar:1,000 μm. (C) Representative immunofluorescence images showed the expression of EGFP and PP2Ac in the ARC of AAV virus injected mice, scale bar: 20 μm; 3V: Third ventricle. (D) Western blot analysis of AKT, pAKT, CREB1, and pCREB1 in the hypothalamus of ARC Control and ARC Ppp2ca mice. (E) The representative LH pulse curves of ARC Control and ARC Ppp2ca mice, # indicates the peak LH concentration. (F) Mean LH, amplitude, peak frequency, peak LH, and basal LH in whole blood of male mice from ARC Control and ARC Ppp2ca mice ( n = 6). (G) Serum T levels of male mice from ARC Control and ARC Ppp2ca mice ( n = 6). (H) Total sperms, motile sperms, progressive sperms, VAP, VCL, VSL, ALH, BCF, LIN, and STR of male mice from ARC Control ( n = 15) and ARC Ppp2ca mice ( n = 13). Data are presented as mean ± SEM, ∗ indicates a significant difference (∗ means p < 0.05, ∗∗ means p < 0.01, ∗∗∗ means p < 0.001, ∗∗∗∗ means p < 0.0001), “ns” indicates non-significant difference, Student’s t test.

Article Snippet: CREB1 Rabbit Polyclonal Antibody , Proteintech , Cat# 12208-1-AP; RRID: AB_2245417.

Techniques: Expressing, Injection, Virus, Fluorescence, Microscopy, Immunofluorescence, Western Blot, Control, Concentration Assay

Journal: iScience

Article Title: The sperm quality in DIO male mice is linked to the NF-κB signaling and Ppp2ca expression in the hypothalamus

doi: 10.1016/j.isci.2025.112110

Figure Lengend Snippet:

Article Snippet: CREB1 Rabbit Polyclonal Antibody , Proteintech , Cat# 12208-1-AP; RRID: AB_2245417.

Techniques: Recombinant, Modification, SYBR Green Assay, Expressing, Virus, Enzyme-linked Immunosorbent Assay, Hormonal Assay, Fluorescence, Staining, Software, RNA Sequencing

Journal: Molecular Medicine

Article Title: Deletion of GPR81 activates CREB/Smad7 pathway and alleviates liver fibrosis in mice

doi: 10.1186/s10020-024-00867-y

Figure Lengend Snippet: Antibodies for immunoblotting and immunohistochemistry staining

Article Snippet: CREB1 Rabbit Polyclonal Antibody , Beyotime Biotechnology , AF6566 , WB 1:1000.

Techniques: Western Blot, Immunohistochemistry

Journal: Molecular Medicine

Article Title: Deletion of GPR81 activates CREB/Smad7 pathway and alleviates liver fibrosis in mice

doi: 10.1186/s10020-024-00867-y

Figure Lengend Snippet: GPR81 modulated CREB/Smad7 pathway. A – D WT mice with CCl 4 -induced liver fibrosis were supplemented with or without DHBA for 8 weeks. A The hepatic contents of cAMP were detected by ELISA kit and were expressed as a fold change relative to the CCl 4 group (n = 4). B The phosphorylation and total protein levels of CREB were determined (n = 4). C The mRNA levels of Smad7 were examined (n = 4). D The protein levels of Smad7, phosphorylated-Smad3 (p-Smad3), and total Smad3 (Smad3) were determined (n = 4). E – H WT mice and GPR81 KO mice were treated with CCl 4 for 8 weeks to induce liver fibrosis. E The hepatic contents of cAMP were detected by ELISA kit and were expressed as a fold change relative to the WT group (n = 4). F The phosphorylation and total protein levels of CREB were determined (n = 4). G The mRNA levels of Smad7 were examined (n = 4). H The protein levels of Smad7, phosphorylated-Smad3 (p-Smad3), and total Smad3 (Smad3) were determined (n = 4). All data were expressed as mean ± SD

Article Snippet: CREB1 Rabbit Polyclonal Antibody , Beyotime Biotechnology , AF6566 , WB 1:1000.

Techniques: Enzyme-linked Immunosorbent Assay, Phospho-proteomics

Journal: Molecular Medicine

Article Title: Deletion of GPR81 activates CREB/Smad7 pathway and alleviates liver fibrosis in mice

doi: 10.1186/s10020-024-00867-y

Figure Lengend Snippet: Treatment with GPR81 activator inhibited CREB/Smad7 pathway and promoted HSCs activation. Human hepatic stellate cells LX-2 with TGF-β1 exposure were supplemented with DHBA for 24 h. A The phosphorylation and total protein levels of CREB were determined (n = 4). B The mRNA levels of Smad7 were examined (n = 4). C The protein levels of Smad7, phosphorylated-Smad3 (p-Smad3), and total Smad3 (Smad3) were determined (n = 4). D The mRNA expressions of COL1A1 were examined (n = 4). E The protein levels of α-SMA and COL1A1 were examined (n = 4). All data were expressed as mean ± SD

Article Snippet: CREB1 Rabbit Polyclonal Antibody , Beyotime Biotechnology , AF6566 , WB 1:1000.

Techniques: Activation Assay, Phospho-proteomics

Journal: Molecular Medicine

Article Title: Deletion of GPR81 activates CREB/Smad7 pathway and alleviates liver fibrosis in mice

doi: 10.1186/s10020-024-00867-y

Figure Lengend Snippet: The schematic diagram of the mechanisms underlying the pro-fibrotic activities of GPR81. The development of liver fibrosis is associated with upregulation of GPR81. GPR81 is a Gi-coupled receptor that decreases the level of cAMP and suppresses the activation of CREB. The suppressed CREB might reduce the expression of Smad7, an inhibitory Smad that competitively inhibits the activation of Smad3. Smad3 is essential for the transcription of collagen I and other pro-fibrotic genes. The reduction of Smad7 might weaken the inhibition of Smad3, resulting in uncontrolled Smad3 signaling, enhanced collagen production, and aggravated liver fibrosis

Article Snippet: CREB1 Rabbit Polyclonal Antibody , Beyotime Biotechnology , AF6566 , WB 1:1000.

Techniques: Activation Assay, Expressing, Inhibition

Journal: Molecular Medicine

Article Title: Deletion of GPR81 activates CREB/Smad7 pathway and alleviates liver fibrosis in mice

doi: 10.1186/s10020-024-00867-y

Figure Lengend Snippet: Antibodies for immunoblotting and immunohistochemistry staining

Article Snippet: Phospho-CREB1 (Ser133) Rabbit Polyclonal Antibody , Beyotime Biotechnology , AF5785 , WB 1:1000.

Techniques: Western Blot, Immunohistochemistry

Journal: iScience

Article Title: TET2 is recruited by CREB to promote Cebpb , Cebpa , and Pparg transcription by facilitating hydroxymethylation during adipocyte differentiation

doi: 10.1016/j.isci.2023.108312

Figure Lengend Snippet:

Article Snippet: Rabbit polyclonal anti-CREB , proteintech , Cat#12208-1-AP; RRID: AB_2245417.

Techniques: Virus, Plasmid Preparation, Recombinant, Transfection, Lysis, CCK-8 Assay, BIA-KA, Cholesterol Assay, CRISPR, Software, Magnetic Beads

Journal: Journal of breast cancer

Article Title: Norepinephrine/β 2 -Adrenergic Receptor Pathway Promotes the Cell Proliferation and Nerve Growth Factor Production in Triple-Negative Breast Cancer.

doi: 10.4048/jbc.2023.26.e25

Figure Lengend Snippet: Figure 5. Norepinephrine/β2-adrenergic receptor signal promotes the growth of triple-negative breast cancer cells via ERK activation. (A) Phosphorylation of ERK and CREB after NE treatment of MD-MB231 cells. After starvation, the cells were treated with 10 µM NE/ISO/Ate/Pro/ICI for 6 hours. Cell lysates were subjected to immunoblotting with antibodies against p-ERK (1:1,000), ERK (1:1,000), p-CREB (1:800), CREB (1:800), and β-actin (1:1,000). Quantitation of the western blotting results is shown in the bar graph. Data from three independent experiments are expressed as means ± SD. (B) Phosphorylation of ERK and CREB after NE treatment of BT549 cells. Data from three independent experiments are expressed as means ± SD. (C) CCK-8 assay of the MD-MB231 and BT549 cells. After starvation, MD-MB231 and BT549 cells were treated with 10 µM NE/ISO/KT5720/U0126 for 24 hours to measure the cell viability. Data are shown as means ± SD from triplicate experiments. NE = norepinephrine; ISO = isoprenaline; Ate = atenolol; Pro = propranolol; SD = standard deviation; CCK = Cell Counting Kit; β2-AR = β2-adrenergic receptor. *p < 0.01, †p < 0.001.

Article Snippet: Anti-tyrosine hydroxylase (TH) Ab (75875), anti-S100 Ab (112), and anti-β2-AR Ab (182136) were purchased from Sigma (St. Louis, USA); anti-CREB Ab (12208-1-AP), phosphorylated anti-CREB Ab (ser133), anti-ERK Ab (16443-1-AP), Phosphorylated anti-ERK Ab (T202/Y204), and anti-NGF Ab (52918) were purchased from Cell Signaling Technology (Danvers, USA); anti-beta-actin Ab was purchased from Beijing Zhongshan Jinqiao Company (Beijing, China); anti-TrkA BAS-bs (10210R) was purchased from Beijing Biosynthesis Biotechnology Co., Ltd. (Beijing, China); horseradish peroxidase (HRP)-conjugated Goat Anti-Rabbit IgG (H+L) (SA00001-2) was purchased from Proteintech Group (Rosemont, USA).

Techniques: Activation Assay, Phospho-proteomics, Western Blot, Quantitation Assay, CCK-8 Assay, Standard Deviation, Cell Counting