Anti-Smt3 antibody

• ELISA

AbReview18914****

ELISA - Anti-Smt3 antibody (AB14405)

ab14405 detecting Smt3 in Escherichia coli recombinant protein (BL21(DE3) by Direct ELISA. Wells were coated with 10µg each of diluted control protein (without SUMO fusion - Negative Controls) and target protein (with SUMO fusion protein) in bicarbonate buffer. Wells were blocked with 5% milk for 1 hour at room temperature. The samples were incubated with primary antibody (1/5000 in TBST +5% milk) for 45 minutes. An HRP-conjugated goat polyclonal to rabbit IgG (1/40000) was used as the secondary antibody.

This image is courtesy of an anonymous Abreview

• WB

Unknown

Western blot - Anti-Smt3 antibody (AB14405)

Western blot of SUMO-GFP fusion proteins cleaved by insect cell protein extracts. Anti-SUMO antibody, generated by immunization with recombinant yeast SUMO, was tested by western blot against several constructs of SUMO-GFP fusion proteins after cleavage by proteases in insect cell protein extracts. These constructs contained various linkers between the SUMO and GFP portion of the fusion proteins.

All lanes:

Western blot - Anti-Smt3 antibody (ab14405) at 1/1000 dilution

Lanes 1, 3, 5, 7, 9, 11 and 13:

Lysate prepared from E.coli expressed Yeast SUMO protein and SUMO-GFP fusion protein after incubation with lysed insect cells at 2 µg

Lanes 2, 4, 6, 8, 10, 12 and 14:

Lysate prepared from SUMO-GFP fusion protein after incubation with lysed insect cells in the presence of 2% SDS at 2 µg

Predicted band size: 12 kDa

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