Protein concentration in the hippocampus was determined as in our previous study. A quantity of 20–40 μg of protein was loaded onto an 8–15% gradient polyacrylamide gel, electrophoretically transferred to a polyvinylidene difluoride membrane and probed with the following primary antibodies: RIP1 antibody (1:1000, Abcam, United States); RIP3 antibody (1:1000, Abcam, United States); neuroligin (1:500, Abcam, United States); neurexin (1:500, Abcam, United States); PSD95 (1:1000, Cell Signaling); CREB (1:1000, Cell Signaling); phospho-CREB (p-CREB) (1:1000, Cell Signaling); BDNF (1:1000, Santa Cruz Biotechnology); β-actin (1:2000, Sigma-Aldrich) was used as an internal control. Secondary antibodies were horseradish peroxidase conjugated to mouse anti-rabbit/mouse immunoglobulin G (1:10,000, Sigma-Aldrich).
Phospho creb p creb
Manufactured by Cell Signaling Technology
Sourced in United States
Phospho-CREB (p-CREB) is a monoclonal antibody product that specifically recognizes the phosphorylated form of the cAMP response element-binding protein (CREB). CREB is a transcription factor that plays a crucial role in cellular processes such as metabolism, cell growth, and neuronal plasticity. The phosphorylation of CREB at specific serine residues is a key regulatory mechanism that modulates its activity and downstream signaling.
Automatically generated - may contain errors
Lab products found in correlation
Psd95, by Cell Signaling Technology (2 mentions)
Brain derived neurotrophic factor (bdnf), by Santa Cruz Biotechnology (1 mentions)
Conjugated to horseradish peroxidase, by Merck Group (1 mentions)
Rip1 antibody, by Abcam (1 mentions)
Rip3 antibody, by Abcam (1 mentions)
Mouse anti rabbit mouse immunoglobulin g, by Merck Group (1 mentions)
β actin, by Merck Group (1 mentions)
α melanocyte stimulating hormone α msh, by Santa Cruz Biotechnology (1 mentions)
Anti rabbit antibody, by Thermo Fisher Scientific (1 mentions)
Ascorbic acid, by Merck Group (1 mentions)
2 2 azino bis, by Merck Group (1 mentions)
β actin, by Santa Cruz Biotechnology (1 mentions)
Quantity one software, by Bio-Rad (1 mentions)
3 protocols using phospho creb p creb
1
Hippocampal Protein Expression Analysis
2
Tyrosinase Activity and Melanogenesis Assay
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2,2′-Azino-bis (3-ethylbenzthiazoline-6-sulfonic acid) (ABTS), 2,2-diphenyl-1-picrylhydrazyl (DPPH), Folin–Ciocalteu reagent (for total phenolics), dimethyl sulfoxide-d6 (DMSO), mushroom tyrosinase (EC 1.14.18.1), and ascorbic acid were obtained from Sigma Aldrich (St. Louis, MO, USA). All reagents used were of analytical grade. Phospho-CREB (p-CREB) and phospho-PKA (p-PKA) were purchased from Cell Signaling (Danvers, MA, USA). Antibodies against tyrosinase (TYR), TRP-1, TRP-2, microphthalmia-associated transcription factor (MITF), CREB, PKA, α-melanocyte-stimulating hormone (α-MSH), and β-actin were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). Horseradish peroxidase-conjugated anti-mouse, anti-goat, and anti-rabbit antibodies were purchased from Invitrogen (Carlsbad, CA, USA).
3
Protein Expression Analysis in Hippocampus
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Western blotting was performed using antibodies specific for postsynaptic density-95 (PSD-95) (catalog number 3409s), cyclic adenosine monophosphate (cAMP)-response element binding protein (CREB) (catalog number 9197), and phospho-CREB (p-CREB) (catalog number 9198s), all obtained from Cell Signaling Technology (Danvers, MA, USA), and β-actin (catalog number AICP001; Sizhengbo, Beijing, People’s Republic of China). Hippocampal tissues were homogenized, centrifuged (12,000 rpm, 4°C, 15 minutes), and the supernatants were collected. Proteins were separated via sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transblotted onto polyvinylidene difluoride membranes. The membranes were blocked and incubated with the relevant primary antibody (1:1,000, 4°C, overnight). Membranes were washed and incubated with horseradish peroxidase-conjugated secondary antibodies (24°C, 1 hour). Immunoreactive proteins were visualized by a chemiluminescence reaction, and data were analyzed using Quantity One software (Bio-Rad Laboratories Inc., Hercules, CA, USA).
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