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AF488 NHS ester

CAS No. : 1374019-99-4

MCE 国际站：AF488 NHS ester

产品活性：AF488 NHS ester 是一种胺特异性荧光探针 (Em=525 nm)。AF488 NHS ester 在水和生物样品中，与巯基和胺反应，并在衍生化后改变它们的化学结构和荧光性质。

研究领域：Others

作用靶点：Fluorescent Dye

In Vitro: IgG fluorescent labeling and anti-IgG solid phase peptide libraries screening of AF488 NHS ester
(1) Dissolve the lyophilized immunoglobulin in a solution of 50 mM sodium phosphate, 20 mM sodium chloride and PH 8.3 at a concentration of 5 g/L.
(2) Dissolve 1 mg of AF488 NHS ester in 100 µL extra dry DMF, add 1 mL of solution in (1), and incubate at room temperature for 1 h.
(3) Collect samples with Amicon Ultra 0.5-mL Centrifugal Filter Unit equipped with 3-kDa MWCO filters.
(4) The deprotected libraries of hexameric or tetrameric are washed in 50 mM sodium phosphate, 150 mM sodium chloride, PH 7.4 (PBS) solution with 5× settled resin volume washing three times to balance.
(5) Dilute IgG-AF488 with 50 mM sodium phosphate, 150 mM sodium chloride, 0.2% Tween (PH 7.4) solution until the final concentration is 1.3 mg/mL.
(6) Incubate (4) and (5) at 2-8°C overnight.
(7) Wash resin beads with 50 mM sodium phosphate, 150 mM sodium chloride, 0.1% Tween 20, PH 7.4 (PBS-T).
(8) The resin is deposited in a 96-well plate of 40 µL PBS-T with one bead per well, and then the fluorescence microscope is used to image at 10× magnification. Under 480 nm excitation, Alexa Fluor 488 fluorescence measurement and fluorescence screening are carried out with 510 nm emission intensity as the threshold.
AF488 NHS ester storage solution
Prepare 20 mM AF488 NHS ester with DMF.
Note: AF488 NHS ester storage solution is recommended to be stored in dark at -20°C after sub-packaging.

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