Western blot analysis was performed on membrane enriched cell extracts of Jurkat (Lane 1) and tissue lysate of Rat Brain (Lane 2). The blot was probed with Anti- Gnb1 Rabbit Polyclonal Antibody (Product# PA1-725, 2ug/ml) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4ug/ml, 1:2500 dilution). A 37 kDa band corresponding to Gnb1 was observed across all lysates tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
|Tested species reactivity||Bovine, Fruit fly, Human, Mouse, Sheep, Rat|
|Published species reactivity||Bovine, Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic Peptide: R(8) Q E A E Q L K N Q I R D A R K A C(25)|
|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 *|
|Western Blot (WB)||1-3 µg/ml|
|Western Blot (WB)||See 5 publications below|
PA1-725 detects transducin beta (T beta) from sheep, bovine, mouse and drosophila.
PA1-725 has been successfully used in Western blot procedures. By Western blot, this antibody detects an ~37 kDa protein representing Tr beta from sheep and bovine retinal and optic nerve extracts as well as mouse eye samples and drosophila.
The PA1-725 immunizing peptide corresponds to amino acid residues 8-25 from human Tr beta. This sequence is completely conserved between human, amphibian, bovine and rat. PA1-725 immunizing peptide (Cat. # PEP-154) is available for use in neutralization and control experiments.
Vision involves the conversion of light into electrochemical signals that are processed by the retina and subsequently sent to and interpreted by the brain. The process of converting light into an electrochemical signal begins when the membrane-bound protein, rhodopsin, absorbs light within the retina. Photoexcitation of rhodopsin causes the cytoplasmic surface of the protein to become catalytically active. In the active state, rhodopsin activates transducin, a GTP binding protein. Once activated, transducin promotes the hydrolysis of cGMP by phosphodiesterase (PDE). The decrease of intracellular cGMP concentration causes the ion channels within the outer segment of the rod or cone to close, thus causing membrane hyperpolarization and, eventually, signal transmission. Rhodopsin activity is believed to be shut off by phosphorylation followed by binding of the soluble protein, arrestin.
Transducin, once activated by rhodopsin, promotes the hydrolysis of cGMP by PDE. The subunit composition of transducin differs between different photoreceptor cells. Rod transducin consists of rod transducin alpha (Tr alpha), T beta, and T gamma. Cone transducin is composed of cone transducin alpha (Tc alpha), T beta and T gamma. Differential transducin subunit composition of transducin is believed to be responsible for the different light sensitivities between photoreceptive cells.
Phosphorylation of phosducin accelerates rod recovery from transducin translocation.
PA1-725 was used in western blot to study the ability of phosducin phosphorylation to markedly accelerate the recovery of rods from transducin translocation following exposure to rod-saturating light
|Belcastro M,Song H,Sinha S,Song C,Mathers PH,Sokolov M||Investigative ophthalmology and visual science (53:3084)||2012|
RAS-converting enzyme 1-mediated endoproteolysis is required for trafficking of rod phosphodiesterase 6 to photoreceptor outer segments.
PA1-725 was used in western blot 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|
Mechanism for the regulation of mammalian cGMP phosphodiesterase6. 1: identification of its inhibitory subunit complexes and their roles.
PA1-725 was used in western blot to study the regulatory mechanism for mammalian cGMP phosphodiesterase 6 actvity
|Yamazaki A,Bondarenko VA,Matsuura I,Tatsumi M,Kurono S,Komori N,Matsumoto H,Hayashi F,Yamazaki RK,Usukura J||Molecular and cellular biochemistry (339:215)||2010|
Disruption of the gene encoding the beta1-subunit of transducin in the Rd4/+ mouse.
PA1-725 was used in western blot to investigate the role of Gnb1 for Rd4 retinal disease
|Kitamura E,Danciger M,Yamashita C,Rao NP,Nusinowitz S,Chang B,Farber DB||Investigative ophthalmology and visual science (47:1293)||2006|
Phototransduction in transgenic mice after targeted deletion of the rod transducin alpha -subunit.
PA1-725 was used in western blot to investigate transducin subunits' functional specificity in phototransduction
|Calvert PD,Krasnoperova NV,Lyubarsky AL,Isayama T,Nicoló M,Kosaras B,Wong G,Gannon KS,Margolskee RF,Sidman RL,Pugh EN,Makino CL,Lem J||Proceedings of the National Academy of Sciences of the United States of America (97:13913)||2000|