EGFP | Enhanced Green Fluorescence Protein
AS13 2700 | clonality: polyclonal | host: rabbit | reactivity: EGFP
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|Recommended dilution||1 : 1000-1 : 5000 (IHC), (WB)|
|Expected | apparent MW|
|Not reactive in|
EGFP has Excitation max (nm): 488; Emission max (nm): 509; Extinction coefficient (Cm-1M-1): 61000.
to be added when available
50 mg of leaves were homogenized in 650 μl extraction buffer (100mM glicine, 10mM EDTA and 100mM NaCl pH 9,5) and diluted with equal volume of Laemli buffer. 2 μl extract of Nicotiana benthamiana leaves infiltrated with GFP-containing construct (1-4), non-infiltrated N. benthamiana (6) and GFP transgenic N. benthamiana were separated on 12% SDS-PAGE and blotted overnight on to PVDF membrane using wet tank transfer. After 1h blocking at room temperature (RT) in 5% milk powder solution blots were incubated with EGFP antibody (AS 13 2700) 1h (RT) at a dilution of 1:10 000 in 1% milk powder (1X PBST). Antibody solution was decanted and the blot was washed 3 times for 5 min in 1X PBST. As a secondary antibody, blot was incubated with goat anti-rabbit IgG HRP conjugated from Agrisera (AS09 602) 1h at a dilution of 1:10 000 in 1XPBST with agitation. Washing was carried out as previously described. Blot was developed with ECL, exposure was made using ChemiDoc equipment in signal accumulation mode (50 sec).
Courtesy of Dr. Ágnes Dalmadi, NAIK, Hungary
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