product information

background
 

Glycogen phosphorylase (EC 2.4.1.1) is an enzyme which is catalyzing the rate limiting step in the degradation of glycogen in animals by releasing glucose-1-phosphate from the terminal alpha1,4-glycosidic bond.

immunogen
 

KLH-conjugated peptide derived from the sequence of human glycogen phosphorylase P11217

antibody format
 

rabbit,

polyclonal

affinity purified serum

lyophilized

quantity
 

200 µg

- for reconstitution add 200 µl of sterile water

storage
 

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

tested applications
 

western blot (WB), immunoprecipitation (IP)

related products
 

AS09 454 | anti-glycogen debranching enzyme

additional information
 

This antibody can detect GP in crude tissue homogenate in both, liver and muscle tissue and is able to immunoprecipiate GP.

application information

recommended dilution
 

1: 2000 (WB), 5 µg (IP)

expected | apparent MW
 

97 kDa

confirmed reactivity
 

human, mouse, rabbit,rat

predicted reactivity
 

bovine, dog, hen,horse,pig, Xenopus laevis, Zebrafish, atlantic salmon, Drosophila melanogaster

not reactive in
 

no confirmed exceptions from predicted reactivity known in the moment

additional information
 

samples used to test this antibody were: muscle and liver homogenates, purified glycogen. This antiboy will recogize glycogen phosphorylase from both liver and muscle. It has been used in ICC on human primary myotubes

selected references
 

Zhu et al. (2011). High post-mortem temperature combined with rapid glycolysis induces phosphorylase denaturation and produces pale and exudative characteristics in broiler Pectoralis major muscles. Meat Science

Parker et al. (2007). AMP-activated protein kinase does not associate with glycogen alpha-particles from rat liver. Biochem. Biophys. Res. Commun. 362:811-815.

application example

(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.

The GP antibody did not immunoprecipitate glycogen phosphorylase from rat liver.

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