Arf1 G-LISA Activation Assay Kit (colorimetric)

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With the new Arf1 G-LISA kit you can now measure Arf1 activation from cell and tissue samples in less than 3 h. G-LISA requires only 1-5% of the material needed for a conventional pull-down assay. You will also be able to handle large sample numbers and generate quantitative results. For a more detailed introduction on G-LISA assays and a listing of other available G-LISA kits, see our main G-LISA page.
The Arf1 G-LISA kit contains an Arf-GTP-binding protein linked to the wells of a 96 well plate. Active, GTP-bound Arf1 in cell/tissue lysates will bind to the wells while inactive GDP-bound Arf1 is removed during washing steps. The bound active Arf1 is detected with a Arf1 specific antibody. The degree of Arf1 activation is determined by comparing readings from activated cell lysates versus non-activated cell lysates.

The kit contains sufficient reagents to perform 96 Arf1 activation assays. Since the Arf-GTP affinity wells are supplied as strips and the strips can be broken into smaller pieces, each kit can be used for anywhere from one to multiple assays. The following components are included in the kit:
96 well Arf1-GTP binding plate (divisible into 12 strips of 8 wells each)
Anti-Arf1 antibody
HRP-labeled secondary antibody
Arf1 control protein (constitutively active Arf1)
Cell Lysis buffer
Wash buffer
Binding buffer
Antigen presenting buffer
Antibody dilution buffer
HRP detection reagent A
HRP detection reagent B
HRP stop solution
Precision Red advanced protein assay reagent (Cat. # ADV02)
Protease inhibitor cocktail (Cat. #PIC02)
Manual with detailed protocols and extensive troubleshooting guide

96-well plate spectrophotometer capable of reading 490 nm wavelength
Multichannel or multidispensing pipettor
Orbital microplate shaker capable of at least 200 rpm shaking (400 rpm is optimal)