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Plant/Algal cell antibodies / Oryza sativa


AMY | Alpha-amylase

Art no: AS10 712
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product information

background  

α-Amylases are hydrolytic enzymes responsible for the mobilization of the starch into metabolizable sugars. This process provides the energy for the growth of roots and shoots and is crucial during germination of cereal seeds.These enzymes are coded by a multigene family and even thought other amylolytic enzyme participate in the process of starch breakdown, the contribution of α-amylase is the prerequisite for the initiation of this process.

immunogen  

KLH-conjugated synthetic peptide derived from known Oryza sativa P17654

antibody format  

rabbit

polyclonal

serum

lyophilized

quantity  

100 µl

for reconstitution add 100 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)

related products   AS10 716 | Ramy 3D | Alpha-amylase isozyme 3D
additional information  

antibody will detect all alpha amylase isoforms from rice, barley and other cereals

application information

recommended dilution  

1 : 5000 with standard ECL (WB)

expected | apparent MW  

43-48 kDa (depending upon the isoform)

confirmed reactivity  

Oryza sativa, Panicum virgatum

predicted reactivity  

cereals

not reactive in  

no confirmed exceptions from predicted reactivity known in the moment

additional information  

to be added when available

selected references  

to be added when available, antibody released in December 2010.


application examples

alpha-Ramy detection in rice using western blot

25 µg of total protein from Oryza sativa seedlings (from 1 day to 5 days of imbibition on filter paper at 28°C) extracted with an SDS Extraction Buffer (60mM Tris-HCl pH 8.0, 2% SDS, 1,5% Sucrose) were separated on XT CRITERION 10%Bis-Tris (BioRad) SDS-PAGE and blotted 1h to PVDF. Blot was blocked immediately in milk in TBS-T for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a diluition of 1: 5,000 in milk in TBS-T for 3h at RT with agitation. The antibody solution was decanted and the blot was rinsed briefly once, then washed once for 15 min and 3 times for 5 min in TBS-T at RT with agitation. Blot was incubated in secondary antibody (goat anti-rabbit IgG HRP conjugated, AS09 602) diluited 1:20 000 in milk in TBS-T for 50 min at RT and then washed as above and developed for 3 min with standard ECL. Images of the blot were obtained using BioSpectrum AC Imaging System (UVP). Exposure time was 30 min. The arrow indicates alpha-amylase (between 47kDa and 50KDa as expected).

Courtesy Valeria Banti and prof. Pierodomenico Perata, PlantLab, Scuola Superiore Sant'Anna


||| For applications or usage on species others than stated above Agrisera offers a payment-after-testing option. To learn more about this or for any questions on this product, please use the LiveChat option in the left menue bar or contact us at  support@agrisera.com



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