showForm Anti-GDE | glycogen debranching enzyme antibodies

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GDE / AGL | Glycogen debranching enzyme

AS09 454 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Human, Mouse, Rabbit, Rat

GDE / AGL | Glycogen debranching enzyme  in the group Antibodies for Human/Animal  / Human Proteins / Other Human proteins at Agrisera AB (Antibodies for research) (AS09 454)

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Product Information

Immunogen

KLH-conjugated peptide derived from the sequence of human glycogen debranching enzyme UniProt: P35573

Host Rabbit
Clonality Polyclonal
Purity Affinity purified serum
Format Lyophilized
Quantity 200 µg
Reconstitution 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 Immunoprecipitation (IP), Western blot (WB)
Recommended dilution 5 µg (IP), 1 : 2000 (WB)
Expected | apparent MW

175 kDa

Reactivity

Confirmed reactivity Human, Mouse, Rabbit, Rat
Predicted reactivity Dog
Not reactive in No confirmed exceptions from predicted reactivity are currently known.

Application examples

Application examples

Application example

western blot using anti-GDE antibodies

(1) rat liver homogenate (50 µg of total protein), (2) GDE IP from 500 µg rat liver homogenate (3) GDE IP from 10 µg purified rat liver glycogen (see Parker et al, BBRC 2007) 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 GDE antibodies (diluted 1/2000) 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 withchemiluminescent detection reagent. Images were detected using the Fuji LAS-3000 system.

To obtain GDE-bound immunoprecipitates, 5 µg GDE antibody was incubated with 500 µg rat liver homogenate or 10 ug purified rat liver glycogen 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. 100µ l, 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.

Additional information

Additional information

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

Antibody will work as an excellent marker for glycogen. In tissues with no glycogen extression there will be no signal using this antibody. 

Related products

Related products

AS09 455 | Anti-glycogen phosphorylase, rabbit antibodies

Secondary antibodies

Background

Background

Synonymes:Glycogen debrancher, 4-alpha-glucanotransferase (EC=2.4.1.25), Oligo-1,4-1,4-glucantransferase, amylo-alpha-1,6-glucosidase, amylo-1,6-glucosidase (EC=3.2.1.33), Dextrin 6-alpha-D-glucosidase,amylo-1, 6-glucosidase, 4-alpha-glucanotransferase

Product citations

Selected references Sottnik et al. (2018). Elucidating the role of Agl in bladder carcinogenesis by generation and characterization of genetically engineered mice. Carcinogenesis. 2018 Nov 7. doi: 10.1093/carcin/bgy139.
Richmond et al. (2018). Glycogen debranching enzyme (AGL) is a novel regulator of non-small cell lung cancer growth. Oncotarget, 2018, Vol. 9, (No. 24), pp: 16718-16730.
Oldenburg et al. (2016). CD44 and RHAMM are essential for rapid growth of bladder cancer driven by loss of Glycogen Debranching Enzyme (AGL). BMC Cancer. 2016 Sep 5;16:713. doi: 10.1186/s12885-016-2756-5.
Pagliarani et al. (2014). Glycogen storage disease type III: A novel Agl knockout mouse model. Biochim Biophys Acta. 2014 Aug 1. pii: S0925-4439(14)00250-6. doi: 10.1016/j.bbadis.2014.07.029.

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