rabbit polyclonal antibody p creb  (Proteintech)

Journal: Heliyon

Article Title: New insights into the downregulation of cytochrome P450 2E1 via nuclear factor κB-dependent pathways in immune-mediated liver injury

doi: 10.1016/j.heliyon.2023.e22641

Figure Lengend Snippet: Effects of PDTC treatment on the expression of PKC, CREB, P-CREB, CYP2E1, NF-κB, and IκBα in immune-mediated liver injury. Rats were administered injections of BCG (125 mg/kg, i.v. on day 1) or BCG (125 mg/kg, i.v. on day 1) + PDTC (25, 50, or 100 mg/kg/d, i.p. on days 11, 12, and 13). Liver proteins were extracted to determine the expression of PKC, CREB, P-CREB, CYP2E1, NF-κB, and IκBα. Equal amounts (30 μg) of protein were subjected to SDS-PAGE followed by Western blot analysis using anti-PKC, anti-CREB, anti-P-CREB, anti-CYP2E1 anti–NF–κB and anti-IκBα antibodies. The results were normalised to the levels of GAPDH and β-tubulin. Western blotting images showing the protein expression of (A) PKC, (B) P-CREB, (C) CREB, (D) CYP2E1, (E) NF-κB, and (F) IκBα in the rat liver. Image Quant software was used to quantify expression. Data are expressed as the mean ± S.D. of three independent experiments (n = 3). The asterisks above the bars indicate differences between groups. Comparisons between groups were performed using one-way ANOVA followed by Tukey's test. Abbreviations: PDTC, ammonium pyrrolidine dithiocarbamate; PKC, protein kinase C; CREB, cAMP-response element binding protein, P-CREB, phospho-CREB; CYP2E1, cytochrome P450 2E1; NF-κB, nuclear factor κB; IκBα, an inhibitor of NF-κB subunit alpha; BCG, Bacillus Calmette–Guerin; SDS-PAGE, sodium dodecyl sulphate-polyacrylamide gel electrophoresis; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; ANOVA, analysis of variance.

Article Snippet: Enzyme-linked immunosorbent assay (ELISA) kits for measuring the levels of rat tumour necrosis factor-alpha (TNF-α), interleukin (IL)-1β, phosphodiesterase (PDE4), and cAMP, BCA protein kit, the sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) preparation kit, as well as rabbit polyclonal antibodies against CYP2E1 (catalogue number: PB0186), PKC (catalogue number: PBM0401), CREB (catalogue number: PB0513), phospho-CREB (P-CREB, catalogue number: P00577), NF-κB (catalogue number: BA0610), glyceraldehyde-3-phosphate dehydrogenase (GAPDH; catalogue number: BA2913), and β-tubulin (catalogue number: A01857-1), were purchased from Wuhan Boster Biological Engineering Co. Ltd. (Hubei, China).

Techniques: Expressing, SDS Page, Western Blot, Software, Binding Assay, Polyacrylamide Gel Electrophoresis

Journal: Scientific reports

Article Title: Standardized Centella asiatica (ECa 233) extract decreased pain hypersensitivity development in a male mouse model of chronic inflammatory temporomandibular disorder.

doi: 10.1038/s41598-023-33769-w

Figure Lengend Snippet: Figure 7. Phosphorylated cAMP response element-binding protein (p-CREB) expression in trigeminal subnucleus caudalis (TNC) in all groups at different time points. (A) An example of p-CREB staining by immunohistochemistry at the ipsilateral TNC at post–CFA-injection day 28, scale bar = 100 µm. (B,C) Expression of p-CREB in both sides of TNCs in different groups. In the ipsilateral TNC at post–CFA-injection days 14, 21, and 28 and in the contralateral TNC at post–CFA-injection day 28, the p-CREB expression in the complete Freund’s adjuvant–treated (CFA) group increased compared with that in the Sham group (on the ipsilateral side: day 14 [F(5,48) = 35.179, P < 0.05], day 21 [F(5,48) = 34.179, P < 0.05], day 28 [F(5,48) = 33.789, P < 0.05]; on the contralateral side: day 28 [F(5,48) = 26.885, P < 0.05]), whereas that in the groups treated with ibuprofen and 100 mg/kg ECa 233 (Ibu and 100ECa groups, respectively) significantly decreased compared with that in the CFA group (day 14 on the ipsilateral side: Ibu [F(5,48) = 21.225, P < 0.05], 100ECa [F(5,48) = 25.247, P < 0.05]; day 21 on the ipsilateral side: Ibu [F(5,48) = 22.375, P < 0.05], 100ECa [F(5,48) = 26.027, P < 0.05]; day 28 on the ipsilateral side: Ibu [F(5,48) = 24.335, P < 0.05], 100ECa [F(5,48) = 26.247, P < 0.05]; day 28 on the contralateral side: Ibu [F(5,48) = 26.245, P < 0.05], 100ECa [F(5,48) = 27.161, P < 0.05]). Both 30ECa and 300ECa showed no difference from the CFA group (day 14 on the ipsilateral side: 30ECa [F(5,48) = 40.485, P = 0.207], 300ECa [F(5,48) = 39.532, P = 0.236]; day 21 on the ipsilateral side: 30ECa [F(5,48) = 40.532, P = 0.196], 300ECa [F(5,48) = 39.532, P = 0.179]; day 28 on the ipsilateral side: 30ECa [F(5,48) = 41.485, P = 0.187], 300ECa [F(5,48) = 39.852, P = 0.196]; day 28 on the contralateral side: 30ECa [F(5,48) = 40.495, P = 0.197], 300ECa [F(5,48) = 39.972, P = 0.194]) (mean ± SEM; n = 3/group at each time; *P < 0.05 compared with the Sham group, #P < 0.05 compared with the CFA group, two-way analysis of variance followed by Fisher’s least significant difference post hoc test).

Article Snippet: The TG paraffin sections were stained with neuroinflammatory markers rabbit monoclonal anti-NaV1.7 antibody (AB5390; Merck KGaA, Darmstadt, Germany) and rabbit polyclonal anti-CGRP antibody (SC-57053; Santa Cruz Biotech, Dallas, TX, USA), whereas the TNC sections were immunolabeled with a rabbit polyclonal anti-p-CREB antibody (SC-81486; Santa Cruz Biotech, Dallas, TX, USA), rabbit polyclonal anti-CGRP antibody (SC-57053; Santa Cruz Biotech, Dallas, TX, USA), and rabbit polyclonal anti-OX42 antibody (SC-52; Santa Cruz Biotech, Dallas, TX, USA) for the expression of activated microglia.

Techniques: Binding Assay, Expressing, Staining, Immunohistochemistry, Injection, Adjuvant

Journal: Scientific reports

Article Title: Standardized Centella asiatica (ECa 233) extract decreased pain hypersensitivity development in a male mouse model of chronic inflammatory temporomandibular disorder.

doi: 10.1038/s41598-023-33769-w

Figure Lengend Snippet: Figure 9. Expressions of CGRP and NaV1.7 in TG, and expressions of CGRP, p-CREB, and activated microglia in TNC in Sham, CFA, and 100ECa groups at different time points. (a) Expressions of CGRP and NaV1.7 in the ipsilateral TG. (b) Expressions of CGRP, p-CREB, and activated microglia in the ipsilateral TNC. (c) Expressions of CGRP and NaV1.7 in the contralateral TG. (d) Expressions of CGRP, p-CREB, and activated microglia in the contralateral TNC. (mean ± SEM; n = 3/group/time point; *P < 0.05 compared with the Sham group, #P < 0.05 compared with the CFA group, two-way analysis of variance followed by Fisher’s least significant difference post hoc test).

Article Snippet: The TG paraffin sections were stained with neuroinflammatory markers rabbit monoclonal anti-NaV1.7 antibody (AB5390; Merck KGaA, Darmstadt, Germany) and rabbit polyclonal anti-CGRP antibody (SC-57053; Santa Cruz Biotech, Dallas, TX, USA), whereas the TNC sections were immunolabeled with a rabbit polyclonal anti-p-CREB antibody (SC-81486; Santa Cruz Biotech, Dallas, TX, USA), rabbit polyclonal anti-CGRP antibody (SC-57053; Santa Cruz Biotech, Dallas, TX, USA), and rabbit polyclonal anti-OX42 antibody (SC-52; Santa Cruz Biotech, Dallas, TX, USA) for the expression of activated microglia.

Techniques:

Journal: Neural Plasticity

Article Title: Electroacupuncture Zusanli (ST36) Relieves Somatic Pain in Colitis Rats by Inhibiting Dorsal Root Ganglion Sympathetic-Sensory Coupling and Neurogenic Inflammation

doi: 10.1155/2023/9303419

Figure Lengend Snippet: Antibodies used for Western blot analysis.

Article Snippet: p-CREB , Rabbit polyclonal , 1/200 , AP0019, ABclonal, Wuhan, China.

Techniques: Western Blot

Journal: PLoS ONE

Article Title: Cytoplasmic Location of α1A Voltage-Gated Calcium Channel C-Terminal Fragment (Ca v 2.1-CTF) Aggregate Is Sufficient to Cause Cell Death

doi: 10.1371/journal.pone.0050121

Figure Lengend Snippet: (A) In a control brain affected with Parkinson's disease, the immunoreactivity against CREB is present but weak and homogeneous in the neuronal cytoplasm of two Purkinje cells. (B) In SCA6 human Purkinje cells, focally accentuated immunoreactive structures (arrows) are seen in the cytoplasm. (C) In ten control brains, a vast majority of Purkinje cells showed weak and diffuse immunoreactivity for the anti-CREB antibody as demonstrated in the . In contrast, aggregate-like CREB-immunoreactive structures were seen in the Purkinje cells from three SCA6 cerebella. Please refer to the Materials and Methods for the detailed description on the criteria of CREB-immunoreactivities. (D) On double immunofluorescence analysis using a rabbit polyclonal CREB antibody and 1C2, a mouse monoclonal antibody against expanded polyQ tracts, microscopic Ca v 2.1 aggregates in the cytoplasm of the SCA6 Purkinje cells indeed co-localized with CREB (arrows). (For A&B: scale bars: 50 µm; for D&E: scale bar: 10 µm). (E) In three SCA6 cerebella, approximately 50% of Purkinje cells containing 1C2-positive polyQ aggregates in the cytoplasm showed co-localization of the polyQ aggregates and CREB. In contrast, the Purkinje cells in control cerebella did not show 1C2-positive aggregates resulting in no co-localizations.

Article Snippet: Either of the following primary antibodies was used; mouse monoclonal anti-c-Myc antibody (diluted with phosphate buffered saline (PBS) into 1∶100) (Santa Cruz Biotechnology, CA, USA), rabbit polyclonal A6RPT-#5803 (1∶500), rabbit polyclonal anti-CREB antibody (1∶500) (Cell Signaling Technology, MA, USA), rabbit polyclonal anti-p-CREB antibody (1∶100) (Cell Signaling Technology), mouse monoclonal anti-expanded 1C2 (1∶1000)(Millipore, CA, USA).

Techniques: Immunofluorescence