NifH | Positive control/quantitation standard

Nitrogenase is involved in biological fixation of nitrogen to assimilable ammonia.This product is a recombinant protein standard, source: Nostoc/Anabaena 7120.

Standard curve: 3 loads are recommended (0.5, 2 and 4 μl) .For most applications a sample load of 0.2 μg of chlorophyll will give a NifH signal in this range.Positive control: a 2 μl load per well is optimal for most chemiluminescent detection systems. This standard is stabilized and ready and does not require heating before loading on the gel.Please note that this product contains 10% glycerol and might appear as liquid but is provided lyophilized. Allow the product several minutes to solubilize after adding water. Mix thoroughly but gently Take extra care to mix thoroughly before each use, as the proteins tend to settle with the more dense layer after freezing.

For reconstitution add 95 µl of sterile water. Please notice that this product contains 10% glycerol and might appear as liquid but is provided lyophilized.

The NifH protein standard can be used in combination with global anti-NifH antibodies to quantitate NifH protein from a wide range of cyanobacterial species. Global antibodies are raised against highly conserved 15 amino acid sequence found in NifH proteins.Quantitative western blot: detailed method description, video tutorial

Levitan et a. (2010) . Regulation of nitrogen metabolism in the marine diazotroph Trichodesmium IMS101 under varying temperatures and atmospheric CO concentrations. Environ. Microbiol (Epub ahead of print)

Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please, remember to spin tubes briefly prior to opening them to avoid any losses that might occur from lyophilized material adhering to the cap or sides of the tubes.