ACT | Actin (monoclonal)
AS16 4111 | Clonality: Monoclonal | Host: Mouse | Reactivity: Arabidopsis thaliana
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Store at -20°C for up to 1 year. 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 liquid material adhering to the cap or sides of the tubes.
1 : 1000 (WB)
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No confirmed exceptions from predicted reactivity are currently known
50-200µg of total protein from: Arabidopsis thaliana (4), Camelina sativa (5) Arabidopsis lyrata (6) extracted with exaction buffer (50mM Tris-HCl pH7.5, 150mM NaCl, 10%(v/v) glycerol, 0.5mM EDTA and 0.5% NP40) and denatured with SDS-sample buffer at 950°C for 5 min, proteins were separated on 10 % SDS-PAGE and blotted 1h to PVDF membrane using semi-dry transfer. Blots were blocked with 5 % milk for 0.5h/RT with agitation. Blot was incubated in the primary antibody at a dilution of 1: 1 000 for ON/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 PBST at RT with agitation. Blot was incubated in Agrisera matching secondary antibody (anti-mouse IgG (H&L), horse radish peroxidase conjugated, AS09 627) diluted to 1: 5 000 in for 1h/RT with agitation. The blot was washed as above, developed for 5 mins with Agrisera ECL Bright and detected with film.
Courtesy Dr Jiaqi Sun, McGill University, Montreal, Canada
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