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ATG9 | Autophagy-related protein 9 (C-terminal)

AS16 4072   | Clonality: Polyclonal  |  Host: Rabbit | Reactivity: Arabidopsis thaliana

ATG9 | Autophagy-related protein 9 (C-terminal) in the group Antibodies for Plant/Algal  / Protein Modifications / Autophagy-related and Ubiquitin-like Proteins at Agrisera AB (Antibodies for research) (AS16 4072)

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Product Information

Immunogen KLH-conjugated peptide derived from Arabidopsis thaliana ATG9 protein sequence, UniProt: Q8RUS5, TAIR: AT2G31260

Host Rabbit
Clonality Polyclonal
Purity Affinity purified serum, in PBS pH 7.4
Format

Lyophilized

Quantity

50 µg

Reconstitution

For reconstitution add 50 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)
Recommended dilution

1 : 10 000 (WB)

Expected | apparent MW

99.5 | 100 kDa

Reactivity

Confirmed reactivity Arabidopsis thaliana
Predicted reactivity Aquilegia coerulea, Brassica napus, Capsella rubella, Daucus carota, Eutrema salsugineum, Gossypium hirsutum, Juglans regia, Lactuca sativa, Noccaea caerulescens, Theobroma cacao, Trema orientale
Not reactive in

Nicotiana tabacum

Application examples

Application examples Application example


Western blot using anti-ATG9 antibodies

Increasing amount of microsomal fraction of Arabidopsis thaliana cell culture (1,2,3).  Microsomal protein was extracted accoring to Maudoux et al. (2000).   and denatured with loading buffer (2 % SDS) at 37°C for 1h before SDS-PAGE gel electrophoresis and blotted to PVDF using semi-dry. Blots were blocked with 3 % nonfat milk + 0.1 % goat serum in TBS-T (Tween 20)  for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 10 000 + 1 % goat serum for ON at 4°C with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, 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 (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1:10 000 in TBS-T + 1 % goat serum for 1h at RT with agitation. The blot was washed as above and developed with chemiluminescent detection reagent. Exposure time was  seconds.
Red marker band has apparent mass of 72 kDa. 

Courtesy of Dr. Henri Batoko, University of Louvain, Belgium 

Additional information

Plant ATG9 is most likely N-glycosylated, and glycosylated membrane proteins can migrate as more than one form in SDS-PAGE (smear); the protein most likely forms functional dimers which could be detected also after western blotting.

Usage of microsomal fraction is highly recommended.

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Background

Background ATG9 (Autophagy-related protein 9) is required for autophagy that plays an essential role in plant nutrient recycling. 
Alternative name: AtAPG9.

Product citations

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