标题：供应 beta－检测试剂盒

迈晨科技  beta－检测试剂盒

-Galactosidase Assay Kit   beta－检测试剂盒

Catalogue：MPK006   Storage: 4－8 ºC

Kit Components:

Kit Components      Chemicals      Size      Cat #

I      Extraction Buffer      10 mL      MC045

II      Assay Buffer (-gal)      60 mL      MC048

III      Substrate Solution (ONPG)      10 mL      MC049

IV      Stop Solution      10 mL      MC007.1

V      pCMV--gal      10 g      MN008

Procedures:

Preparation of Cell Lysates

      Remove the media from the tissue culture plate wells.

      Add 100 μl Extraction Buffer into each well to cover the cells.

      Incubate the plate on shaker platform at room temperature for 5-10 minutes.

      Place the plates on ice and proceed to the next step.

Preparation of Reaction Solution

Calculate how much Reaction Solution you need (200l per well of 96-well plate). Below is the table for preparation of 100 assays.

Materials      Quantity     

Note: The kit contains sufficient reagents for 250 assays performed in 96-well tissue culture plate.

*-Mercaptoethanol was not provided in the kit

Assay Buffer      20 mL      

ONPG      4 mL     

-Mercaptoethanol*      100 L     

      For each well, add 200 l assay buffer and 5-10 l of cell lysates.

      Incubation at 37 ºC for 20-60 minutes till yellow shade is observed.

      Add 100 l stop solution (1 M Na2CO3).

      Measure O.D. at 420 nm.

      Calculate the units of -galactosidase activity using the formula: -gal (unit) = (O.D.420 nm X 380 X 10) / (minutes of incubation X volume (l) of cell extract)

Reference:

Herbomel et al. Cell 39:653, 1984

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