rabbit anti creb1 polyclonal antibody  (Proteintech)

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: 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

Journal: Frontiers in Pharmacology

Article Title: Effects of Chang-Kang-Fang Formula on the Microbiota-Gut-Brain Axis in Rats With Irritable Bowel Syndrome

doi: 10.3389/fphar.2022.778032

Figure Lengend Snippet: Primer sequences for rat target genes.

Article Snippet: Next, the sections were incubated with the anti-5-HT1A rabbit polyclonal antibody (Cat. No.D260012), anti PKA rabbit polyclonal antibody (Cat. NO.D164496), anti-CREB1 (Phospho-Ser133) rabbit polyclonal antibody (Cat. No.D155349), and anti-BDNF polyclonal antibody (Cat. No.D221057) (1:50) (Sangon Biotech, China), overnight at 4°C in a dilution ratio of 1:100 using the BondTM Primary Antibody Diluent (Servicebio, China).

Techniques: Sequencing

Journal: Frontiers in Pharmacology

Article Title: Effects of Chang-Kang-Fang Formula on the Microbiota-Gut-Brain Axis in Rats With Irritable Bowel Syndrome

doi: 10.3389/fphar.2022.778032

Figure Lengend Snippet: Chang-Kang-Fang formula affects the levels of 5-HT, 5-HT1AR, BDNF, PKA, and CREB in IBS rats. (A,B) 5-HT levels in colon and hippocampus. (C,D) The relative expression of 5-HT1AR in colon and hippocampus. (E,F) The relative expression of BDNF in colon and hippocampus. (G,H) The relative expression of BDNF in colon and hippocampus. (I,J) The relative expression of PKA in colon and hippocampus. (K,L) The relative expression of CREB in colon and hippocampus. Values are presented as the means ± SD ( n = 6). ## p < 0.01, ### p < 0.001 compared to Normal, * p < 0.05, ** p < 0.01 and *** p < 0.001 compared to IBS.

Article Snippet: Next, the sections were incubated with the anti-5-HT1A rabbit polyclonal antibody (Cat. No.D260012), anti PKA rabbit polyclonal antibody (Cat. NO.D164496), anti-CREB1 (Phospho-Ser133) rabbit polyclonal antibody (Cat. No.D155349), and anti-BDNF polyclonal antibody (Cat. No.D221057) (1:50) (Sangon Biotech, China), overnight at 4°C in a dilution ratio of 1:100 using the BondTM Primary Antibody Diluent (Servicebio, China).

Techniques: Expressing

Journal: Frontiers in Pharmacology

Article Title: Effects of Chang-Kang-Fang Formula on the Microbiota-Gut-Brain Axis in Rats With Irritable Bowel Syndrome

doi: 10.3389/fphar.2022.778032

Figure Lengend Snippet: Immunohistochemical staining for the colon and hippocampal (A,B) PKA and (C,D) CREB. Values are presented as the means ± SD ( n = 3). ### p < 0.001 as compared to Normal,* p < 0.05, ** p < 0.01 and *** p < 0.001 as compared to IBS.

Article Snippet: Next, the sections were incubated with the anti-5-HT1A rabbit polyclonal antibody (Cat. No.D260012), anti PKA rabbit polyclonal antibody (Cat. NO.D164496), anti-CREB1 (Phospho-Ser133) rabbit polyclonal antibody (Cat. No.D155349), and anti-BDNF polyclonal antibody (Cat. No.D221057) (1:50) (Sangon Biotech, China), overnight at 4°C in a dilution ratio of 1:100 using the BondTM Primary Antibody Diluent (Servicebio, China).

Techniques: Immunohistochemical staining, Staining