BC2-tag

AS20 4446 | Clonality: Polyclonal | Host: Rabbit | Reactivity: BC2-tag

Product Information

Immunogen KLH-conjugated BC2 tag PDRKAAVSHWQQ derived from beta catenin.

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 deionized 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 dilution 1 : 5000 (WB)

Reactivity

Confirmed reactivity Protein of interest overexpressed wth BC2-tag

Application examples

Application examples Western blot using anti-BC2-tag antibodies

Samples:
10 µg A. thaliana – Wt Col-0 (Negative control) (1)
20 µg A. thaliana – Wt Col-0 (Negative control) (2)
10 µg of A. thaliana expressing POI-BC2t fusion (3)
20 µg of A. thaliana expressing POI-BC2t fusion (4)

10 + 20 µg/well of total protein were extracted from Arabidopsis thaliana leaf material in diluted HENS (25mM HEPES pH 7.7, 1mM EDTA, 2.5 % SDS) and stored at -80°C. Samples were denatured in 1x protein loading dye (0.5% Sodium dodecyl Sulfate, 0.002% Bromophenol Blue, 10% glycerol, and 50 mM Tris-HCl pH6.8) at 95°C for 5 min. Samples were separated on 12% SDS-PAGE gel and blotted 1h to a PVDF membrane (pore size of 0.2 µm), using a semi-dry transfer. Blot was blocked with 10% milk in TBS-T o/n at 4°C without agitation. Blot was incubated in the primary antibody at a dilution of 1:5000 in 10% milk in TBS-T, at RT, for 1h with agitation. The antibody solution was decanted and the blot was rinsed briefly, then washed three times for 10 min in TBS-T at RT with agitation. Blot was incubated in Agrisera matching secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, AS09 602) diluted to 1:25 000 in 10% milk in TBS-T at RT for 1h with agitation. The blot was washed as above and developed for 2 min with ECL substrate. Exposure time was 11 minutes and 2 seconds.

Courtesy of Dr. Patrick Treffon, Elizabeth Vierling Lab Department of Biochemistry and Molecular Biology University of Massachusetts Amherst, USA