FUM1 + FUM2 | Fumarase 1+2 (chloroplastic+mitochondrial)
AS16 3966 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana
|Datasheet||Product citations||Protocols||Add review|
for reconstitution add 50 µl of sterile water
49.9 | 55 kDa
Total protein extracted freshly from Arabidopsis thaliana 14-old-day seedlings grown on ½ MS without sugar in extraction buffer [100 mM NaCl; 50 mM Tris–HCl pH 7.5; 0.5% (v/v) Triton X-100; 1 mM DTT; 1× Complete Protease Inhibitor Cocktail Tablet (Roche, Bassel, Switzerland)] and denatured in Laemeli buffer at 55°C for 5 min. 50 µg of protein extraccts: Arabidopsis thaliana wt (1), fum2-1 mutant (2), fum2-1 + GFP-FUM2 N1 (3), fum2-1 + GFP-FUM2 N2 (4), fum2-1 + GFP-FUM2 N3 (5), fum2-1 + GFP-FUM2 N3 (6), no protein -GFP, fum2-2 (8) were loaded and separated on 12% SDS-PAGE and blotted 1h to PVDF membranes in a semi-dry transfer system. The membrane was blocked with 3% milk for 1h at RT with agitation and then incubated in the primary antibody (anti-FUM) at a dilution of 1: 2 000 for 1h at RT with agitation in TBS-T and milk 3%. The antibody solution was decanted and the blot was rinsed briefly three times in TBS-T for 5 min. Blot was incubated in Agrisera matching secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated AS09 602) diluted to 1:5 000 in for 1h at RT with agitation. The blot was washed as above and developed for 10 min using chemiluminescnce detection reagent in a Gel Doc XR+ System (Bio-rad; Hercules; USA).
fum1 mutant is lethal therefore a band of ca. 60 kda is still present in fum2-1 and fum2-2 background.
Courtesy of Dr. Antoni Garcia Molina, Ludwig-Maximilians-Universität München (LMU), Germany
To be added when available, antibody released in October 2018.
Related products: FUM1 + FUM2 | Fumarase 1+2 (chloroplastic+mitochondrial)
For an extra 20 EUR on your order you will receive:
- Matching s...