GP | glycogen phosphorylase

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AS09 455 | clonality:polyclonal | host:rabbit | reactivity:chicken, human, mouse, rabbit, rat, T.vaginalis G3


54 st
Item No:
AS09 455

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product information

Glycogen phosphorylase (EC 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.


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

Host Rabbit
Clonality Polyclonal
Purity Affinity purified serum
Format Lyophilized
Quantity 50 µg
Reconstitution For reconstitution add 50 ĩl of sterile water

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

Secondary antibodies

Additional information

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

Glycogen phosphorylase is a good marker for determining the degree of postmortem protein denaturation in muscle.  When glycogen phosphorylase denatures it shifts from the sarcoplasmic fraction of the muscle to the myofibrillar fraction of the muscle.  This shift is detectable using SDS-PAGE and western blotting techniques.  The antibody is being used to detect glycogen phosphorylase levels in both sarcoplasmic and myofibrillar protein extracts from chicken muscle.

application information
Recommended dilution

1: 2000 (WB), 5 µg (IP)

Expected | apparent MW

97 kDa

Confirmed reactivity

human, mouse, rabbit,rat, Trichomonas vaginalis G3

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 Dandanell et al. (2016). Maintaining a clinical weight loss after intensive lifestyle intervention is the key to cardiometabolic health. pii: S1871-403X(16)30107-7. doi: 10.1016/j.orcp.2016.09.009.
Bowker and Zhuang (2015). Relationship between water-holding capacity and protein denaturation in broiler breast meat. Poult Sci. 2015 May 25. pii: pev120.
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 Scienc

application example

western blot using anti-GP antibodies

(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.

western blot using anti-glycogen phosphorylase antibodies used on T.vaginalis

The antibody reacts weakly with NCBI GI:121879937 (glycogen/starch/alpha-glucan phosphorylases family protein from T. vaginalis G3 strain) and well with NCBI GI:121911693 (glycogen phosphorylase from T. vaginalis G3 strain). The two TvGPs are 75% identical to one another. 300 nanograms of purified recombinant TvGP (lanes 1 and 2; labels indicate GI number);  10 micrograms of rat skeletal muscle (3)extracted with phosphate-buffered saline, pH 7.4, containing 5 mM 2-ME, 1 mM EDTA, 1% Triton X-100, and 1% protease inhibitor cocktail (Sigma-Aldrich P8340) (lane 3); and 5 microliters of Bio-Rad pre-stained markers (lane 4). All samples were mixed 1:1 with electrophoresis sample buffer (125 mM Tris, pH 6.8, 20% glycerol, 4% SDS, 10% 2-ME, 0.1% bromophenol blue) and solubilized by boiling for 5 min in a waterbath before electrophoresis through a 12 % SDS-PAGE mini-gel. Proteins were blotted 1h to PVDF by tank transfer in buffer comprising 25 mM Tris, 192 mM glycine, and 20% methanol, pH 8.3. Blots were blocked with 5% non-fat dry milk (NFDM) in Tris-buffered saline (TBS; 20 mM Tris, 150 mM NaCl, pH 7.5) for 1h at room temperature (RT) with agitation. Blot was incubated ON with agitation in the primary antibody diluted 1: 2 000 in TBS containing 0.05% Tween 20 (TTBS) and 1% NFDM. The antibody solution was decanted and the blot was rinsed briefly twice, then washed 3 times for 10 min in TTBS at RT with agitation. Blot was incubated for 2h at RT with agitation in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, from Agrisera  (AS09 602) diluted to 1:5 000 in TTBS-1% NFDM. The blot was washed in TTBS as above, rinsed a final time in TBS to remove the Tween 20, and developed with TMB colorimetric substrate (Kirkegaard & Perry Labs, #50-77-00) for about 10 min.

Courtesy of Dr. Melissa Stuart, Microbiology/Immunology Kirksville College of Osteopathic Medicine, USA

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