Anti-EIN2 | Ethylene insensitive 2

• Immunogen: KLH-conjugated peptide chosen from EIN2 of Arabidopsis thaliana, UniProt: Q9S814, TAIR: AT5G03280Chosen peptide is not to be found in EIN3.
• Host: Rabbit
• Clonality: Polyclonal
• Purity: Immunogen 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 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 dilutions: 1 : 2000 (WB)
• Expected | apparent MW: 140 kDa

Application example



40-60 µg of membrane protein (measured with Millipore Direct Detect spec.) isolated following membrane extraction buffer protocol from Dong et al. 2008 (Plant Journal 53(2): 275-286) from 4 day old dark grown Arabidopsis thaliana seedlings (1,3 WT, 2,4 mutant), grown on MS plated with ACC or AVG in the plate, extracted with (10 mM Tris-HCl pH 7.5, 150 mM NaCl, 1 mM EDTA, 10 % glycerol, 1 % Triton X-100, and protease ihibitors cocktail from Sigma) were separated 4-20% Bio-Rad precast SDS-PAGE gel and blotted 1h to PVDF membrane using semi-dry transfer. Blots were blocked with 5% milk for 2h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 2 000 overnight at 4°C with agitation. The antibody solution was decanted and the blot was then washed 5 times for 10 min in TBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, from Agrisera AS09 602) diluted to 1:20 000 in  for 1h at RT with agitation. The blot was washed as above and developed for 5 min with Bio-Rad ECL according to the manufacturer's instructions. Exposure time was 25 min (to see full length EIN2).

Antibody is suitable for detection of full length EIN2. Bands below 140 kDa are non-specific.

Courtesy of Jennifer Marie Shemansky, University of Maryland, USA