Immunofluorescence analysis of CREB was done on 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.25% Triton™ X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with CREB Mouse Monoclonal Antibody (350900) at 2µg/mL in 1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa Flour 488 Rabbit Anti-Mouse IgG Secondary Antibody (A11059) at a dilution of 1:400 for 30 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor 594 Phalloidin (A12381). Panel d is a merged image showing nuclear localization. Panel e shows no primary antibody control. The images were captured at 20X magnification.
|Tested species reactivity||Human, Mouse, Rat|
|Published species reactivity||Rat, Human, Not Applicable|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Recombinant human CREB-A protein|
|Storage buffer||PBS, pH 7.4|
|Contains||0.1% sodium azide|
|Tested Applications||Dilution *|
|ELISA (ELISA)||Assay Dependent|
|Immunocytochemistry (ICC)||1-3 ug/ml|
|Immunofluorescence (IF)||1-3 ug/ml|
|Immunohistochemistry (IHC)||Assay Dependent|
|Western Blot (WB)||1-3 ug/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
Cyclic AMP response element binding protein (CREB) is a basic/leucine zipper transcription factor that binds the cyclic AMP response element (CRE) and activates transcription in response to a variety of extracellular signals including neurotransmitters, hormones, membrane depolarization, and growth and neurotrophic factors. Activation of CREB is dependent upon the phosphorylation of serine 133. Phosphorylation occurs via p44/42 MAP kinase and p90RSK and also via p38 MAP kinase and MSK1. Although CREB will bind DNA independent of its phosphorylation state, only the phosphorylated form is competent as a transcription factor. CREB binding protein (CBP), a transcriptional coactivator that directly interacts with CREB, binds to CREB in the region of serine 133. CREB appears to play an important role in learning and memory.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Long-term treatment with standardized extract of Ginkgo biloba L. enhances the conditioned suppression of licking in rats by the modulation of neuronal and glial cell function in the dorsal hippocampus and central amygdala.
35-0900 was used in immunohistochemistry to assess the effects of chronic treatment with Ginkgo biloba extract on the conditioned emotional response.
|Oliveira DR,Sanada PF,Filho AC,Conceição GM,Cerutti JM,Cerutti SM||Neuroscience (235:70)||2013|
Neuromodulatory property of standardized extract Ginkgo biloba L. (EGb 761) on memory: behavioral and molecular evidence.
35-0900 was used in immunohistochemistry to assess the effects of acute or subacute treatments with Ginkgo biloba leaf extract on rats submitted to a conditioned emotional response
|Oliveira DR,Sanada PF,Saragossa Filho AC,Innocenti LR,Oler G,Cerutti JM,Cerutti SM||Brain research (1269:68)||2009|
|Human||Not Cited||Inactivation of CREB mediated gene transcription by HDAC8 bound protein phosphatase.||Gao J,Siddoway B,Huang Q,Xia H||Biochemical and biophysical research communications (379:1)||2009|
|Not Applicable||0.5 µg/ml||
Schoenheimer effect explained--feedback regulation of cholesterol synthesis in mice mediated by Insig proteins.
35-0900 was used in western blot to uses knockout mice to study the feedback inhibition of cholesterol synthesis in vivo
|Engelking LJ,Liang G,Hammer RE,Takaishi K,Kuriyama H,Evers BM,Li WP,Horton JD,Goldstein JL,Brown MS||The Journal of clinical investigation (115:2489)||2005|