Anti pcreb

Manufactured by Santa Cruz Biotechnology

Sourced in United States

Anti-pCREB is a primary antibody that specifically recognizes the phosphorylated form of the cAMP Response Element Binding (CREB) protein. CREB is a transcription factor that plays a crucial role in various cellular processes, and its phosphorylation is a key regulatory mechanism. The Anti-pCREB antibody can be used to detect and quantify the levels of phosphorylated CREB in biological samples, enabling researchers to study the activation of this important signaling pathway.

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Goat anti-pCREB (2 citations)

3 protocols using anti pcreb

PTEN, CREB, AKT Pathway Analysis

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For Western blotting, immunoblotting was performed with anti-PTEN (Cell Signaling), anti-pCREB, CREB, pAKT, AKT (Santa Cruz Biotech), and PI3K (Cell Signaling) primary antibodies. Secondary antibodies were specific for peroxidase-conjugated anti-mouse IgG or anti-rabbit IgG (Sigma-Aldrich) as needed. The blots were visualized using an enhanced chemiluminescence detection system (Amersham). The samples were normalized to GAPDH (Cell Signaling) and densitometric analysis for protein quantification was evaluated with “Image J” software.

Lai Y.J., Hsu H.H., Chang G.J., Lin S.H., Chen W.J., Huang C.C, & Pang J.H. (2017). Prostaglandin E1 Attenuates Pulmonary Artery Remodeling by Activating Phosphorylation of CREB and the PTEN Signaling Pathway. Scientific Reports, 7, 9974.

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Protein Extraction and Western Blot Analysis

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Protein was extracted from cells or tissues using T-per tissue protein extraction reagent (catalog #78510) lysis buffer. Concentration was measured using the Bradford Assay reagent. 10-40 ug of protein was loaded on SDS-polyacrylamide gel and transferred to a nitrocellulose membrane. The membrane was blotted with anti-Mettl3 (#ab195352 Abcam, #MA5-27527 Invitrogen), anti-Mettl14 (#HPA038002, Sigma), anti-GFP (#GFP-1020 Aves Lab), anti-HA (#3724, Cell Signaling), anti-c-Myc (#ab185656 Abcam, #OP10-200ug Sigma), anti-Avpr2 (#AB1797P, EMD Millipore), anti-Yap1 (#4912, Cell Signaling), anti-pCREB (#9198S, pCreb1), anti-PCNA (#sc-7907, Santa Cruz), or anti-puromycin (#PMY-2A4, Developmental Studies Hybridoma Bank) antibodies. All the primary antibodies were used at 1:1000 dilution, except for anti-puromycin, which was used at 1:50 dilution. Goat anti-rabbit-HRP conjugate (#7074S, Cell Signaling) or anti-mouse HRP-conjugated IgG (#7076S, Cell Signaling) was used as the secondary antibody. HRP-conjugated Actin antibody (#A3865, Sigma) was used at 1:40,000 dilution for measuring total protein. The blots were developed using the X-ray film developer or the Bio-rad digital imager. The protein bands were quantified using the Imagelab software from Bio-rad. Each western blot was repeated at least three times.

Ramalingam H., Kashyap S., Cobo-Stark P., Flaten A., Chang C.M., Hajarnis S., Hein K.Z., Lika J., Warner G.M., Espindola-Netto J.M., Kumar A., Kanchwala M., Xing C., Chini E.N, & Patel V. (2021). A methionine-Mettl3-N6-methyladenosine axis promotes polycystic kidney disease. Cell metabolism, 33(6), 1234-1247.e7.

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Hippocampal Protein Expression Analysis

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Hippocampus homogenate was lysed, centrifuged, and separated via SDS‐PAGE. Total proteins were transferred to PVDF membrane (Amersham). The membrane was blocked in 5% skim milk at 25°C for 1 hr, and then treated with anti‐WNT2, anti‐CREB, anti‐p‐CREB, anti‐BDNF, and anti‐β‐actin (Santa Cruz Biotechnology, US) at 4°C for overnight. Then, the membrane was treated with horseradish peroxidase‐labeled secondary antibody (Invitrogen, US). Chemiluminescence kit was used to visualize the signals (Biobyt, UK).

Liu Z., Yang J., Fang Q., Shao H., Yang D., Sun J, & Gao L. (2021). MiRNA‐199a‐5p targets WNT2 to regulate depression through the CREB/BDNF signaling in hippocampal neuron. Brain and Behavior, 11(8), e02107.

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