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AMY | Alpha-amylase

AS10 712  | Clonality: Polyclonal  |  Host: Rabbit  |  Reactivity: Oryza sativa

AMY | Alpha-amylase  in the group Antibodies Plant/Algal  / Carbohydrates at Agrisera AB (Antibodies for research) (AS10 712)
AMY | Alpha-amylase



DATA SHEET IN PDF

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Product Information

Immunogen

KLH-conjugated synthetic peptide derived from known Oryza sativa P17654

Host Rabbit
Clonality Polyclonal
Purity Serum
Format Lyophilized
Quantity 100 µl
Reconstitution 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 the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.
Tested applications Western blot (WB)
Recommended dilution 1 : 5000 (WB)
Expected | apparent MW

43-48 kDa (depending upon the isoform)

Reactivity

Confirmed reactivity Oryza sativa, Panicum virgatum
Predicted reactivity

Cereals, Hordeum vulgare, Kalanchoe laxiflora


Species of your interest not listed? Contact us
Not reactive in

Pinus strobus

Application examples

Application examples

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 chemiluminescent detection. 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

Additional information

Additional information Antibody will detect all alpha amylase isoforms from rice, barley and other cereals

Related products

Background

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.

Product citations

Selected references Ye et al. (2018). Natural variation in the promoter of rice calcineurin B-like protein10 (OsCBL10) affects flooding tolerance during seed germination among rice subspecies. Plant J. 2018 May;94(4):612-625. doi: 10.1111/tpj.13881.
immunogen:

KLH-conjugated synthetic peptide derived from known Oryza sativa P17654

Reconstitution: For reconstitution add 100 µl of sterile water
Host: Rabbit
Clonality: Polyclonal
Purity: Serum
Format: Lyophilized
Quantity: 100 µl
storage: Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please remember to spin the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.
tested applications: Western blot (WB)
recommended dilution: 1 : 5000 (WB)
calculated | apparent molecular mass [kDa]:

43-48 kDa (depending upon the isoform)

Confirmed reactivity: Oryza sativa, Panicum virgatum
predicted reactivity:

Cereals, Hordeum vulgare, Kalanchoe laxiflora


Species of your interest not listed? Contact us
not reactive in:

Pinus strobus

Picture (footer):

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 chemiluminescent detection. 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

additional information: Antibody will detect all alpha amylase isoforms from rice, barley and other cereals
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.

All references: Ye et al. (2018). Natural variation in the promoter of rice calcineurin B-like protein10 (OsCBL10) affects flooding tolerance during seed germination among rice subspecies. Plant J. 2018 May;94(4):612-625. doi: 10.1111/tpj.13881.

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