Sufficient primary antibody (10-fold diluted) was added to double the volume of the settled mass of embryos followed by a rocking motion for 30 min and then incubated overnight without rocking at 4°.
The protoplasts squeezed from the wild B. hypnoides were added to double volume ice-cold extraction buffer (400 mM sucrose, 50 mM Tris, 20 mM EDTA, 0.2% BSA, 0.2% β-mercaptoethanol, pH 7.8) and then filtered through four layers of cheese cloth.
SalI/SmaI adaptors (Takara) were added to double stranded cDNA which was digested with NotI and size fractionated using a cDNA Size Fractionation Column to remove small cDNA (< 500 bp) (Invitrogen).
Following Yoldas method, titanium isopropoxide was added to double-distilled water.
0.1 M TEAAc (pH 7.0): 1.0 M stock solution was made up by adding to double-distilled water (100 ml), freshly distilled TEA (27.8 ml) and acetic acid (11.4 ml).
A mixture of equal volumes of CAV1 and cytokeratin antibodies was incubated overnight at 4°C, rinsed and added to double-stain polymer adjusted for both mouse (HRP) and rabbit (AP) antibodies (Innovex Biosciences, Pinoles, CA, USA) for 30 min at RT.
Add to a double boiler (bain marie) - water should be in the bottom layer of the double boiler.
I would add to the double standards the endless, irritating prattle on Kim Clijsters's re-emergence after becoming a mother.
The net result is the same as if H and −OH add to the double bond with a regioselectivity opposite to the Markovnikov rule.
A single alkene molecule, called a monomer, can add to the double bond of another to give a product, called a dimer, having twice the molecular weight.
