Immunofluorescence analysis of PDE6 beta was performed using 70% confluent log phase MCF-7 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 PDE6B Rabbit Polyclonal Antibody (PA1722) at 1:250 dilution 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 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic and nuclear localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse|
|Published species reactivity||Bovine, Mouse, Human, Not Applicable|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic Peptides: H(20) Q Y F G (K/R) K L S P E N V A G A C(36)|
|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 *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunohistochemistry (IHC)||Assay dependent|
|Western Blot (WB)||2 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.
PA1-722 detects phosphodiesterase 6 beta (PDE6 beta) from mouse retinal extracts.
PA1-722 has been successfully used in Western blot procedures. By Western blot, this antibody detects an ~90 kDa protein representing PDE6 beta from mouse retinal extracts.
The PA1-722 immunizing peptides correspond to amino acid residues 20-36 from mouse and bovine PDE6 beta protein. These peptides (Cat. # PEP-183) are available for use in neutralization and control experiments.
The second messengers cAMP and cGMP are key regulatory molecules that are involved in a wide variety of signal transduction pathways. Levels of cAMP and cGMP are regulated by their rate of synthesis by nucleotide cyclases and by their rate of hydrolysis by cyclic nucleotide phosphodiesterases (PDEs). PDEs form a superfamily of enzymes that catalyze the conversion of 3-prime, 5-prime-cyclic nucleotides to the corresponding nucleoside 5-prime-monophosphates. PDE6 is the effector enzyme in the G protein-mediated signal transduction cascade in the visual system. There are five different subunits consisting of rod and cone specific catalytic subunits: alpha and quote; (Cone), alpha (Rod), and beta (Rod), the inhibitory subunit gamma, and subunit delta of unknown function (which likely interacts with many other proteins besides the PDE6 family). The catalytic core of the PDE6 system is comprised of alpha and quote;/alpha and quote; homodimers in the cone and alpha/beta heterodimers in the rod. The C-terminus of both the catalytic and inhibitory subunits is modified by methylation, myristyolation and prenylation which have been shown to be critical for proper complex assembly and membrane association.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Halting progressive neurodegeneration in advanced retinitis pigmentosa.
PA1-722 was used in western blot to assess the therapeutic time window to treat retinal degenerative disease with gene therapy
|Koch SF,Tsai YT,Duong JK,Wu WH,Hsu CW,Wu WP,Bonet-Ponce L,Lin CS,Tsang SH||The Journal of clinical investigation (125:3704)||2015|
Gene therapy restores vision in rd1 mice after removal of a confounding mutation in Gpr179.
PA1-722 was used in immunohistochemistry - frozen section to investigate how the Grp179 mutation contributes to bipolar cell-mediated b-waves in mice.
|Nishiguchi KM,Carvalho LS,Rizzi M,Powell K,Holthaus SM,Azam SA,Duran Y,Ribeiro J,Luhmann UF,Bainbridge JW,Smith AJ,Ali RR||Nature communications (6:null)||2015|
Generation of three-dimensional retinal tissue with functional photoreceptors from human iPSCs.
PA1-722 was used in immunocytochemistry to characterize functional photoreceptors from human iPSCs generated from three-dimensional retinal tissue
|Zhong X,Gutierrez C,Xue T,Hampton C,Vergara MN,Cao LH,Peters A,Park TS,Zambidis ET,Meyer JS,Gamm DM,Yau KW,Canto-Soler MV||Nature communications (5:null)||2014|
Mammalian sperm phosphodiesterases and their involvement in receptor-mediated cell signaling important for capacitation.
PA1-722 was used in immunocytochemistry to investigate the expression and function of different PDE isoforms in mature mouse spermatozoa
|Baxendale RW,Fraser LR||Molecular reproduction and development (71:495)||2005|
Function of the asparagine 74 residue of the inhibitory ¿-subunit of retinal rod cGMP-phophodiesterase (PDE) in vivo.
PA1-722 was used in western blot to investigate the effect of N74A mutation on the function of PDE6 in vivo
|Tsang SH,Woodruff ML,Hsu CW,Naumann MC,Cilluffo M,Tosi J,Lin CS||Cellular signalling (23:1584)||2011|
Ccdc66 null mutation causes retinal degeneration and dysfunction.
PA1-722 was used in western blot to identify the role of Ccdc66 in retinitis pigmentosa
|Gerding WM,Schreiber S,Schulte-Middelmann T,de Castro Marques A,Atorf J,Akkad DA,Dekomien G,Kremers J,Dermietzel R,Gal A,Rülicke T,Ibrahim S,Epplen JT,Petrasch-Parwez E||Human molecular genetics (20:3620)||2011|
Functional rescue of degenerating photoreceptors in mice homozygous for a hypomorphic cGMP phosphodiesterase 6 b allele (Pde6bH620Q).
PA1-722 was used in western blot to characterize the phenotype of mice with homozygous missense Pde6b(H620Q) mutations and to study a gene therapy protocol.
|Davis RJ,Tosi J,Janisch KM,Kasanuki JM,Wang NK,Kong J,Tsui I,Cilluffo M,Woodruff ML,Fain GL,Lin CS,Tsang SH||Investigative ophthalmology and visual science (49:5067)||2008|
Phototransduction in a transgenic mouse model of Nougaret night blindness.
PA1-722 was used in western blot to investigate the mechanism for Nougaret night blindness.
|Moussaif M,Rubin WW,Kerov V,Reh R,Chen D,Lem J,Chen CK,Hurley JB,Burns ME,Artemyev NO||The Journal of neuroscience : the official journal of the Society for Neuroscience (26:6863)||2006|
The inhibitory gamma subunit of the rod cGMP phosphodiesterase binds the catalytic subunits in an extended linear structure.
PA1-722 was used in western blot to study the interaction between the inhibitory gamma subunit and the catalytic subunit of the rod cGMP phosphodiesterase.
|Guo LW,Muradov H,Hajipour AR,Sievert MK,Artemyev NO,Ruoho AE||The Journal of biological chemistry (281:15412)||2006|
Transducin activation state controls its light-dependent translocation in rod photoreceptors.
PA1-722 was used in western blot to investigate the role of the GTP-hydrolysis on Gtalpha for light-dependent translocation in rod photoreceptors
|Kerov V,Chen D,Moussaif M,Chen YJ,Chen CK,Artemyev NO||The Journal of biological chemistry (280:41069)||2005|
RAS-converting enzyme 1-mediated endoproteolysis is required for trafficking of rod phosphodiesterase 6 to photoreceptor outer segments.
PA1-722 was used in immunohistochemistry to investigate the role of RAS-converting enzyme 1 RCE1 in intracellular PDE6 transport and photoreceptor viability
|Christiansen JR,Kolandaivelu S,Bergo MO,Ramamurthy V||Proceedings of the National Academy of Sciences of the United States of America (108:8862)||2011|
PARP1 gene knock-out increases resistance to retinal degeneration without affecting retinal function.
PA1-722 was used in immunohistochemistry to study the roles of the PARP1 during retinal degeneration
|Sahaboglu A,Tanimoto N,Kaur J,Sancho-Pelluz J,Huber G,Fahl E,Arango-Gonzalez B,Zrenner E,Ekström P,Löwenheim H,Seeliger M,Paquet-Durand F||PloS one (5:null)||2010|