Tetra·His Antibody, BSA-free (100 µg)

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Use of 1/1500 dilution

Protocol tips
Use of 1/1500 dilution

Publication protocol

Expression of recombinant p12I was assessed in RAc- NPV-HTLVp12I-infected Sf-21 cells as previously de- scribed (Arp et al., 1993). Concomitant infections with wild-type AcNPV were performed, and cells were har- vested in a similar manner. Total protein in the resulting cell lysates was determined using the Bradford reagent (Bio-Rad, Hercules, CA), and 50 􏱌g of each lysate was run on a 15% SDS–PAGE gel containing 8 M urea. After electrophoresis, the gel was either silver-stained or Western blotted (Arp et al., 1993). Blots were blocked for 3 h at room temperature with a blocking reagent (Boehr- inger Mannheim, Laval, Quebec, Canada) diluted in ma- leic acid-buffered saline. The primary antibody, a mono- clonal mouse anti-tetra-His antibody (Qiagen), was applied at a 1/1500 dilution for 12 h at 4°C. After the blots were washed and incubated with the goat anti-mouse secondary antibody conjugated with horseradish perox- idase (Jackson ImmunoResearch Laboratories Inc., West Grove, PA) they were exposed to Western Blot Chemilu- minescence Reagent Plus (NEN Life Sciences, Boston, MA), according to the manufacturer’s specifications, and exposed to Cronex 4 film (Sterling Diagnostic Imaging, Inc., Newark, DE).

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Manufacturer protocol

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