mouse anti plcβ  (Santa Cruz Biotechnology)

Journal: Frontiers in Neuroanatomy

Article Title: Up-regulation of CB 1 cannabinoid receptors located at glutamatergic terminals in the medial prefrontal cortex of the obese Zucker rat

doi: 10.3389/fnana.2022.1004702

Figure Lengend Snippet: Primary antibodies used.

Article Snippet: CB 1 cannabinoid receptor, G i/o α-subunits, and phospholipase C-β 1 (PLC-β 1 ) were detected by immunoblotting with the following antibodies: goat polyclonal anti-CB 1 receptor (CB1-Go-Af450; Frontier Science Co. Ltd, Hokkaido, Japan), rabbit polyclonal anti-Gα o (sc-387; Santa Cruz Biotechnology, Santa Cruz, CA, USA), rabbit anti-Gα i 1 (sc-391; Santa Cruz Biotechnology), rabbit polyclonal anti-Gα i 2 (sc-7276; Santa Cruz Biotechnology), rabbit polyclonal anti-Gα i 3 (sc-262; Santa Cruz Biotechnology) and mouse monoclonal anti-PLC-β 1 (BD Transduction Laboratories, San Diego, CA, USA) ( ).

Techniques: Affinity Purification, Transduction

Journal: Cell & Bioscience

Article Title: The mechanism of Annexin A1 to modulate TRPV1 and nociception in dorsal root ganglion neurons

doi: 10.1186/s13578-021-00679-1

Figure Lengend Snippet: ANXA1 mimics Ac2-26 increases intracellular Ca 2+ , and activates PLCβ via FPR2. a Representative images of the calcium-dependent fluorescence 30, 60 and 120 s after application with scramble (3.3 μM), Ac2-26 (3.3 μM) and Boc2 (10 μM) + Ac2-26 (3.3 μM) in cultured AnxA1 −/− DRG neurons. Green color indicates basal intracellular Ca 2+ concentration (i.e. before depolarization) in Fura-2 dye loaded DRG neurons. Warmer colors (yellow, orange and red) indicate that the cytoplasmic Ca 2+ concentration is relatively higher. Scale bar, 100 μm. b Representative traces showing the time course of changes in ratio (340/380) of calcium-dependent fluorescence during application of scramble (3.3 μM), Ac2-26 (3.3 μM) and Boc2 (10 μM) + Ac2-26 (3.3 μM). c , Quantification of the intracellular Ca 2+ concentration ([Ca 2+ ]) responding to scramble (3.3 μM), Ac2-26 (3.3 μM) and Boc2 (10 μM) + Ac2-26 (3.3 μM) (Ac2-26 versus scramble group, *** P < 0.001; Ac2-26 versus Boc2 + Ac2-26 group, **P < 0.01, one-way ANOVA, post hoc Student’s t test. n = 8 in scramble group, n = 9 in Ac2-26 group and n = 8 in Boc2 + Ac2-26 group). d Representative protein band images of the pPLCβ and PLCβ after application with scramble (3.3 μM), Ac2-26 (3.3 μM) and Boc2 (10 μM) + Ac2-26 (3.3 μM) in cultured AnxA1 −/− DRG neurons. β-actin was used as the internal reference protein. e Quantification of the relative protein intensities of pPLCβ among scramble, Ac2-26 and Boc2 + Ac2-26 groups. The data of relative intensities in scramble group were normalized to 1 (Ac2-26 versus scramble group, *** P < 0.001; Ac2-26 versus Boc2 + Ac2-26 group, *** P < 0.001, one-way ANOVA, post hoc Tukey’s multiple comparisons test, n = 5 in each group). All data are represented as means ± SD

Article Snippet: Then, incubating the membranes with primary antibodies (rabbit polyclonal anti-ANXA1 (sc‐11387, 1:200; Santa Cruz Biotechnology, Santa Cruz, CA, USA), mouse monoclonal antibody against FPR2 (1:1000, Cat#H00002358-M02, Abnova), polyclonal rabbit anti-TRPV1 (1:1000, Cat#ACC-030, Alomone Labs), rabbit polyclonal antibody against TRPA1 (1:2000, Cat#OST00061W, Invitrogen), rabbit polyclonal antibody against TRPM8 (1:1000, Cat#OSR00077W, Invitrogen), mouse monoclonal antibody against Calmodulin (1:1000, Cat#05-173, Sigma-Aldrich), mouse monoclonal anti-PLCβ (1:500, Cat#05-164, Sigma-Aldrich) and rabbit polyclonal anti-pPLCβ (1:1000, #2481, Cell Signaling)) at 4 °C overnight.

Techniques: Fluorescence, Cell Culture, Concentration Assay

Journal: Scientific Reports

Article Title: β 2 -adrenoreceptor Inverse Agonist Down-regulates Muscarine Cholinergic Subtype-3 Receptor and Its Downstream Signal Pathways in Airway Smooth Muscle Cells in vitro

doi: 10.1038/srep39905

Figure Lengend Snippet: ( A ) Cropped gel of PLCβ 1 protein levels in rat ASMCs determined by Western blotting. Rat ASMCs were randomly divided into control group, ICI118,551 1 h and 24 h groups, and received different treatments as described previously. ( B ) The densitometry results of PLCβ 1 protein normalized to a β-actin control. Data were presented as means ± SEM from three independent experiments. DMEM + FBS served as control. There were no significant differences among the three group (P > 0.05). ASMCs, airway smooth muscle cells; DMEM, Dulbecco’s modified Eagle’s medium; FBS, fetal bovine serum; PLCβ 1 , phospholipase Cβ 1 ; SEM, standard error of mean.

Article Snippet: A mouse polyclonal anti-rat anti-PLCβ 1 antibody (cat. no. 610924; 1:1,000) was purchased from Becton Dickinson (Dublin, Ireland).

Techniques: Western Blot, Modification

Journal: Scientific Reports

Article Title: β 2 -adrenoreceptor Inverse Agonist Down-regulates Muscarine Cholinergic Subtype-3 Receptor and Its Downstream Signal Pathways in Airway Smooth Muscle Cells in vitro

doi: 10.1038/srep39905

Figure Lengend Snippet: Rat ASMCs were randomly divided into five groups. Cells were incubated with 10 −5 mM ICI118,551, 10 −5 mM formoterol, 10 −4 mM budesonide, 10 −4 mM budesonide + 10 −5 mM formoterol and control group for 24 h. The M 3 R, PLCβ 1 and IP 3 levels were determined after 15 min of Ach (10 −4 mM) stimulation. Cropped gel of M 3 R ( A ) and PLCβ 1 ( C ) protein levels in rat ASMCs were determined by Western blotting. The densitometry results of M 3 R ( B ) or PLCβ 1 ( D ) were normalized to β-actin control. The expressions of IP 3 were evaluated by ELISA ( E ). Data were presented as means ± SEM from three independent experiments. DMEM + FBS served as control. * Significant difference as compared with 24 h of ICI118,551 treatment alone (P < 0.05). Ach, acetylcholine; ASMCs, airway smooth muscle cells; B, budesonide; F, formoterol; DMEM, Dulbecco’s modified Eagle’s medium; FBS, fetal bovine serum; ICI, ICI118,551; IP3, 1,4,5-trisphosphate; M 3 R, muscarine cholinergic subtype-3 receptor; PLCβ 1 , phospholipase Cβ 1 ; SEM, standard error of mean.

Article Snippet: A mouse polyclonal anti-rat anti-PLCβ 1 antibody (cat. no. 610924; 1:1,000) was purchased from Becton Dickinson (Dublin, Ireland).

Techniques: Incubation, Western Blot, Enzyme-linked Immunosorbent Assay, Modification