Immunofluorescent analysis of CREB Binding Protein (green) showing staining in the nucleus of MCF-7 cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a CREB Binding Protein polyclonal antibody (Product # PA1-847) in 3% BSA-PBS at a dilution of 1:100 and incubated overnight at 4°C in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. Actin was stained using Alexa Fluor 554 (red) and nuclei were stained with Hoechst or DAPI (blue). Images were taken at a magnification of 60x.
|Tested species reactivity||Human, Mouse|
|Published species reactivity||Rat, Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to residues A(162) T S S P A T S Q T G P G I C(176) in the nuclear factor binding domain of human CBP.|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|ChIP assay (ChIP)||1-3 ul|
|Immunohistochemistry (Paraffin) (IHC (P))||1:1000-1:5000|
|Western Blot (WB)||1-2 µg/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.
PA1-847 detects cyclic AMP-responsive enhancer binding protein (CREB) binding protein (CBP) from human and mouse tissues and cells.
PA1-847 has been successfully used in ChIP, ICC/IF, Western blot and IHC-P procedures. By Western blot, this antibody detects an ~265 kDa protein representing CBP and a larger, unknown protein at ~400 kDa from HeLa cell lysate.
The PA1-847 immunogen is a synthetic peptide corresponding to residues A(162) T S S P A T S Q T G P G I C(176) in the nuclear factor binding domain of human CBP. The PA1-847 immunizing peptide (Cat. # PEP-052) is available for use in neutralization and control experiments.
Cyclic AMP-responsive enhancer binding protein (CREB) binding protein (CBP) and p300 are closely related transcriptional coactivators that have been shown to directly interact with many different DNA-binding transcription factors including nuclear hormone receptors, CREB (cyclic AMP-responsive enhancer binding protein), c-Fos, c-Jun/v-Jun, c-Myb/v-Myb, TFIIB and MyoD. Both CBP and p300 have been shown to display histone acetyltransferase (HAT) activity, capable of acetylating all four core histone particles in nucleosomes. As a result of HAT activity, it has been suggested CBP and p300 may play a direct role in activating chromatin for transcription.
Single point mutations in CBP have been proposed as causative factors in the developmental abnormalities of Rubinstein-Taybi syndrome (RTS). Although both CBP and p300 appear to function similarly, the inability of p300 to rescue CBP malfunction in RTS suggests intrinsic functional differences between CBP and p300.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Nuclear receptor coactivators function in estrogen receptor- and progestin receptor-dependent aspects of sexual behavior in female rats.
PA1-847 was used in western blot to study the ER and PR action regulated by SRC-1 and CBP in the rat brain
|Molenda-Figueira HA,Williams CA,Griffin AL,Rutledge EM,Blaustein JD,Tetel MJ||Hormones and behavior (50:383)||2006|
The transcriptional coactivators p300 and CBP are histone acetyltransferases.
PA1-847 was used in western blot to investigate the functional properties of CREB binding protein and p300
|Ogryzko VV,Schiltz RL,Russanova V,Howard BH,Nakatani Y||Cell (87:953)||1996|
Nuclear receptor coactivators modulate hormone-dependent gene expression in brain and female reproductive behavior in rats.
PA1-847 was used in immunocytochemistry to demonstrate the regulatory role of nuclear receptors in ER transcriptional activity in brain and the expression of hormone-dependent behavior.
|Molenda HA,Griffin AL,Auger AP,McCarthy MM,Tetel MJ||Endocrinology (143:436)||2002|