U-box domain-containing protein 12

AS13 2728 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Oryza sativa

Product Information

Immunogen

KLH-conjugated synthetic peptide derived from Oryza sativa OsPUB12 protein, UniProt: Q5VRH9

Host Rabbit

Clonality Polyclonal

Purity Serum

Format Lyophilized

Quantity 50 µl

Reconstitution For reconstitution add 50 µl of sterile water

Storage The antibody may be stored at -20℃ for one year in its original formulation. Additionally, antibody may be stored at 2℃ to 8℃ for up to 1 month without detectable loss of activity. Avoid repeated freeze-thaw cycles of the diluted antibody.

Store at 4°C; make aliquots to avoid working with a stock. Please, remember to spin tubes briefly prior to opening them to avoid any losses that might occur from liquid material adhering to the cap or sides of the tubes.

Tested applications Western blot (WB)

Recommended dilution 1 : 1000 (WB)

Expected | apparent MW

66 | 66 kDa

Reactivity

Confirmed reactivity Oryza sativa

Not reactive in No confirmed exceptions from predicted reactivity are currently known.

Application examples

Application examples Application example:

Total protein from Oryza sativa rice (CV. 9311) flag leaf at the tillering stage was ground into a fine powder in liquid nitrogen. An 800 ul aliquot of extraction buffer: 62.5 mM TRIS-HCl (pH 7.4), 10% glycerol, 0.1% SDS, 2 mM EDTA, 1 mM phenylmethylsulphonyl fluoride (PMSF), 5% (v/v) b-mercaptoethanol, was added to each 300 mg powder sample. The mixture was vortexed and then chilled on ice for 10 min. Samples were centrifuged at 12,000 rpm for 10 min. at 4℃, and the supernatant was collected and stored at –70℃. The protein concentrations of the rice samples were determined using the Bradford method (Bradford, 1976). 20 µg of protein was separated on 12 % SDS-PAGE and blotted 1h to PVDF.
Blots were blocked with for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 1 000 for 1h at RT 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 for 1h at RT with agitation. The blot was washed as above and developed for 5 min with ECL according to the manufacturer’s instructions. Exposure time was 30 sec.