Glycogen Phosphorylase (GP) Antikörper

Antigen: Glycogen Phosphorylase (GP)

Klonalität: Polyklonal

Wirt: Kaninchen

Reaktivität: Human, Maus, Kaninchen, Ratte (Rattus)

Applikation: Western Blot (WB), Immunpräzipitation (IP)

Produktnummer: ABIN334639

Produktbeschreibung

Immunogen: KLH-conjugated peptide derived from the sequence of humanglycogen phosphorylase P11217

Format: Lyophilized

Beschreibung: Glycogen phosphorylase (EC 2.4.1.1) is an enzyme which is catalyzing the ratelimiting step in the degradation of glycogen in animals by releasingglucose-1-phosphate from the terminal alpha1,4-glycosidic bond.

Spezifität: Predicted reactivity: bovine, dog, hen,horse,pig, Xenopus laevis, Zebrafish, atlantic salmon,Drosophila melanogaster.

Anwendungen

Applikationshinweise: Recommended Dilution 1: 2000 (WB), 5 µg (IP). Additional Information: This antibody can detect GP in crude tissue homogenate in both, liver and muscletissue and is able to immunoprecipiate GP.

Reinheit: Affinity purified serum

Lagerung: store lyophilized/reconstituted at -20°C, once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please, remember to spin tubes briefly prior toopening them to avoid any losses that might occur from lyophilized materialadhering to the cap or sides of the tubes.

Forschungsgebiet: Enzyme

Beschränkungen: Nur für Forschungszwecke einsetzbar

Bilder

(1) rat liver homogenate (50 µg of total protein), (2) 10 ug purified rat liver glycogen (see Parker et al, BBRC 2007), (3) Human skeletal muscle homogenate (50 µg of total protein), (4) GP IP from 500 ug Human skeletal muscle homogenate were resuspended into SDS PAGE sample buffer, boiled, electrophoresed on a pre-cast 4-15% gradient gel (Invitrogen, CA, USA) and transferred to PVDF membrane. Following blocking in 5% milk / PBST, membranes were probed with GP antibodies (diluted 1/1000) for 1h at room temperature with rocking, followed by washing in 3 times for 5 min in PBS-T at room temperature with agitation. Blots were incubated in secondary antibody (Protein-G coupled to horse radish peroxidase conjugated – Bio-Rad) diluted 1:3000 for 1h at room temperature with agitation. The blots were washed as above and developed for 5 min with ECL (Invitrogen, CA, USA). Images were detected using the Fuji LAS-3000 system. To obtain GP-bound immunoprecipitates, 5 ug GP antibody was incubated with 500 ug human skeletal muscle homogenate together with 1 ml IP wash buffer (50 mM Hepes pH 7.5, 150 mM NaCl, 10% glycerol, 1% NP-40, 1 mM EGTA, 10 mM NaPPi and 100 mM NaF) and incubated at 4°C with mixing for 90 minutes. 100 ul, 20% Protein-A Sepharose was then added for a further 30 minutes at 4°C with mixing. Immunoprecipitates were collected by centrifugation at 7,000 rpm for 1 minutes and washed three times with IP wash buffer followed by resuspension into SDS PAGE sample buffer, boiling and electrophoresis as above. NB: The GP antibody did not immunoprecipitate glycogen phosphorylase from rat liver