Immunofluorescence analysis of CaM Kinase IV was performed using 70% confluent log phase SH-SY5Y cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with CaM Kinase IV Rabbit Polyclonal Antibody (PA1542) at 2ug/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (A27034) at a dilution of 1:2000 for 45 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® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Dog, Human, Mouse, Non-human primate, Plant, Rat, Xenopus|
|Published species reactivity||Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic Peptide: V(127) E K G Y Y S E R D A A D A V K Q(143) C|
|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 *|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Immunohistochemistry (Paraffin) (IHC (P))||1:20-1:100|
|Western Blot (WB)||0.5 - 1 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.
|Western Blot (WB)||See 1 publications below|
PA1-542 detects CaM kinase IV (CaMKIV) from human, mouse, rat, canine, primate, arabidopsis and Xenopus Oocyte samples.
PA1-542 has been successfully used in Western blot and immunohistochemistry procedures. By Western blot, this antibody detects an ~55 kDa protein representing CaMKIV in HeLa cell lysate.
PA1-542 immunizing peptide corresponds to amino acid residues 127-143 from mouse CaMKIV. This sequence is 94% conserved in rat and human CaMKIV. This peptide (Cat. PEP-109) is available for use in neutralization and control experiments.
CaM kinase IV (CaMKIV), in a calcium and calmodulin dependent manner, is implicated in phosphorylating numerous transcription factors including the CCAAT enhancer binding protein (C/EBP), serum response factor (SRF), and cyclic AMP response element binding protein (CBP). The nuclear localization of this protein is consistent with its role in mediating calcium-dependent gene expression. CaMKIV is particularly abundant in testis, T-cells, and neurons but is also found in other tissues to varying degrees. In neurons, CaMKIV is thought to play an important role in synaptic plasticity via its gene regulatory effects. In T-cells, this protein plays an important role in calcium signaling which could affect the transcription regulatory protein, nuclear factor of activated T-cells (NFAT). CaMKIV is encoded, along with calspermin, by the CaMKIV gene. It has been found that, in testes, CaMKIV is expressed in germ cells and found to be associated with chromatin. The association of CaMKIV with chromatin suggests a potential role in chromatin remodeling during nuclear condensation in spermatids.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Potential role of cAMP response element-binding protein in ethanol-induced N-methyl-D-aspartate receptor 2B subunit gene transcription in fetal mouse cortical cells.
PA1-542 was used in western blot to investigate the intracellular signaling pathways related to NR2B gene transcription regulated by long-term ethanol treatment
|Rani CS,Qiang M,Ticku MK||Molecular pharmacology (67:2126)||2005|