Creb 9197s

Manufactured by Cell Signaling Technology

Sourced in United States

CREB (9197s) is a primary antibody that recognizes the CREB (cAMP Response Element Binding Protein) protein. CREB is a transcription factor that plays a crucial role in the regulation of gene expression in response to a variety of cellular signals, including cAMP, calcium, and growth factors.

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5 protocols using creb 9197s

Antibody-based Protein Expression Analysis

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The antibodies were used as follows: mouse monoclonal antibody to glyceraldehyde-3-phosphate dehydrogenase (GAPDH, MAB374, Merck, 1:10000); rabbit polyclonal phosphorylated form of cAMP-dependent protein kinase (pPKA (Thr197), 44988A, Thermo Fisher Scientific, 1:1000); rabbit polyclonal to the recombinant fragment corresponding to a region within amino acids 1 and 351 of PKA catalytic subunit alpha (PA5-21842, Invitrogen, 1:1000); mouse monoclonal to phosphorylated form of cAMP response element-binding protein (CREB (Ser 133), 9196S, Cell Signaling, 1:500); rabbit monoclonal to cAMP response element-binding protein (CREB, 9197S, Cell Signaling, 1:1000); rabbit polyclonal to adenylate cyclase 3 (AC3, PA1-31191, Invitrogen, 1:250); rabbit polyclonal to adenylate cyclase 7 (AC7, PA5-103390, Thermo Fisher Scientific, 1:500); mouse monoclonal to A-kinase anchoring protein 9 (AKAP9, ab32679, Abcam, 1:125); goat monoclonal to A-kinase anchoring protein 9 (AKAP9, ab31307, Abcam, 1:200); rabbit monoclonal to cAMP-specific 3′,5′-cyclic phosphodiesterase 4D (PDE4D, ab171750, Abcam, 1:1000); HRP rabbit polyclonal to β-tubulin (ab21058, Abcam, 1:10000); rabbit polyclonal to MVI (25–6791, Proteus, 1:500); goat anti-mouse IgG antibody, HRP conjugate (AP308P, Millipore, 1:10000); goat anti-rabbit IgG antibody, HRP conjugate (AP307P, Millipore, 1:10000).

Lehka L., Wojton D., Topolewska M., Chumak V., Majewski Ł, & Rędowicz M.J. (2022). Loss of Unconventional Myosin VI Affects cAMP/PKA Signaling in Hindlimb Skeletal Muscle in an Age-Dependent Manner. Frontiers in Physiology, 13, 933963.

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Western Blotting Analysis of Signaling Proteins

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Western blotting was conducted according to a protocol described elsewhere (19 (link)). Proteins of interest were investigated with the primary antibodies to p-AMPK (ab133448), AMPK (ab207442) from Abcam (Cambridge, UK), p-Akt (Thr-308, 13038), Akt (9272), p-IRS-1 (Ser1101, 2385S), p-IRS-1 (Ser302, 2384S), IRS-1 (2382), p-CREB (9198S) and CREB (9197S) from the Cell Signaling Technology (Boston, USA). The signal was detected with chemiluminescent detection system (E412-01-AA, Western Lightning ECL; Vazyme). The signal was collected with a CCD camera system (Light-Capture; ATTO, Tokyo, Japan) and the statistical analysis was conducted with the ImageJ program.

Cao X., Ye X., Zhang S., Wang L., Xu Y., Peng S., Zhou Y., Peng Y., Li J., Zhang X., Han X., Huang W.Y., Jia W, & Ye J. (2021). ADP Induces Blood Glucose Through Direct and Indirect Mechanisms in Promotion of Hepatic Gluconeogenesis by Elevation of NADH. Frontiers in Endocrinology, 12, 663530.

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MITF and CREB Protein Analysis

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Cells were lysed in RIPA buffer (1% NP-40, 150 mM NaCl, 10 mM Tris-HCl at pH 8.0, 1 mM EDTA) with a complete protease inhibitor (Sigma-Aldrich). The proteins were separated by SDS-polyacrylamide gel and transferred to PVDF membranes (Millipore, Billerica, MA). The antibody against MITF was purchased from Abcam (ab12039, Cambridge, UK), those for pCREB (9191s) and CREB (9197s) were purchased from Cell Signalling Technology (Danvers, MA), and the antibodies for tyrosinase (sc-7833) and p53 (sc-126) were purchased from Santa Cruz Biotechnology (Dallas, TX).

Yoon J.E., Kim Y., Kwon S., Kim M., Kim Y.H., Kim J.H., Park T.J, & Kang H.Y. (2018). Senescent fibroblasts drive ageing pigmentation: A potential therapeutic target for senile lentigo. Theranostics, 8(17), 4620-4632.

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Western Blot Protein Expression Analysis

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Detection of protein expression by western blot was carried out according to the established protocols described previously 14 (link). Western blot was performed using the following antibodies: SOX2 (sc-20088) and β-actin (sc-47778) antibodies were purchased from Santa Cruz Biotechnology (Dallas, Texas, USA). OCT4 (ab19857), NANOG (ab80892) and PFKFB4 (ab137785) antibodies were purchased from Abcam (Cambridge, UK). CD44 (3570), Flag (8146s) and CREB (9197s) antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA).

Gao R., Li D., Xun J., Zhou W., Li J., Wang J., Liu C., Li X., Shen W., Qiao H., Stupack D.G, & Luo N. (2018). CD44ICD promotes breast cancer stemness via PFKFB4-mediated glucose metabolism. Theranostics, 8(22), 6248-6262.

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Melanogenesis regulation by molecular pathways

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Dulbecco’s modified Eagle’s medium (DMEM) and fetal bovine serum (FBS) were purchased from WelGene (Gyeongsan-si, Daegu, Korea). Kojic acid, l-tyrosine, l-DOPA, mushroom tyrosinase (EC 1.14.18.1), α-melanocyte-stimulating hormone (α-MSH), and 3-isobutyl-1-methylxanthine (IBMX) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Antibodies against tyrosinase (sc-7833), MITF (sc-11002), phospho-CREB (p-CREB) (sc-81486), p-PKA (sc-12905), PKA (sc-365615), p-JNK (sc-6254), p-ERK (sc-7383), ERK (sc-514302), p-p38 (sc-7973), p38 (sc-535), TFIIB (sc-271736), and β-actin (sc-47778) were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). CREB (#9197s) and JNK (#9252s) antibody purchased from Cell Signaling Technology (Danvers, MA, USA). Peroxidase-labeled anti-rabbit, anti-mouse, and anti-goat immunoglobulins were acquired from GeneTex (Irvine, CA, USA). Reagent-grade chemicals and solvents were purchased from commercial sources. Ultra-pure water was used throughout the experiment.

Jung H.J., Choi D.C., Noh S.G., Choi H., Choi I., Ryu I.Y., Chung H.Y, & Moon H.R. (2021). New Benzimidazothiazolone Derivatives as Tyrosinase Inhibitors with Potential Anti-Melanogenesis and Reactive Oxygen Species Scavenging Activities. Antioxidants, 10(7), 1078.

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