AS21 4532 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana
AS21 4532 Anti-AOX2 | Plant alternative oxidase 2
Immunogen: KLH-conjugated synthetic peptide derived from Arabidopsis thaliana AOX2 protein sequence, UniProt: O22049 , TAIR: At5g64210 Host: Rabbit Clonality: Polyclonal Purity: Antigen affinity purified serum, in PBS pH 7.4 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 : 1000 (WB) Expected | apparent MW: 40.1 | 37.8 kDa (due to N-terminal processing) Confirmed reactivity: Arabidopsis thaliana Predicted reactivity: Species of your interest not listed? Contact us Not reactive in: No confirmed exceptions from predicted reactivity are currently known Samples:
Total protein 1-4
Cytoplasmic fraction
5-8 Nuclear fraction 9-1220 µg/well of total protein extracted freshly from fractionation of Arabidopsis thaliana 7-day-old green seedlings and denatured with 2x Laemmli Sample Buffer at 90°C for 5 min were separated on 12 % SDS-PAGE and blotted 1h to nitrocellulose (pore size of 0.45 um), using wet transfer. Blot was blocked with 5% milk for 1h/RT with agitation. Blot was incubated in the primary antibody at a dilution of 1: 1 000 for ON/4°C with agitation in TBS-T. 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 Agrisera matching secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1:25 000 in for 1h/RT with agitation. The blot was washed as above and developed for 5 min with Agrisera ECLBright. Exposure time was 200 seconds.
Courtesy of Dr. Zhengyao Shao, The University of Texas at Austin , USAAlternative oxidases (AOX) are quinol oxidases located in the inner mitochondrial membrane of plants. They function as terminal oxidases in the alternate electron transport pathway, oxidizing ubiquinone to reduce oxygen to water.