Protocol for western blot needle assay

grow cells: 1. grow cells from stock plates in 1ml LB w/AC to saturation 2. induce with arabinose for 5hours in a total volume of 500ul (1:10 dilution of cells) 3. add 100ul of cells into a 96well plate, triplicates 4. take OD600 5. spin cells down (5500rpm for 5minutes) and flick out liquid

incubate cells with espA: 6. resuspend cells in 100ul of espA with TRIS (5ul espA and 95ul TRIS) 7. incubate for 4hours at 37C with no shaking 8. spin cells down and flick out liquid

lyse cells with lysis buffer

spin down the cell junk

Run an SDS gel to isolate the espA protein

Protein blot on nitrocellulose

Label with specific antibody (that is conjugated to something like alkaline phosphatase)

detect antibody

reference: western blot method