NifH | Positive control/quantitation standard

(Cat#: AS01 021S)
AS01_021S
AS01_021S_1
Description
  • Format: Lyophilized
  • Quantity: 100 µl
  • Reconstitution: For reconstitution add 90 µl of sterile water. Please notice that this product contains 10% glycerol and might appear as liquid but is provided lyophilized.
  • Storage: 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.
  • Tested applications: Western blot (WB)
  • Recommended dilutions: 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.
  • Expected | apparent MW: 34 kDa (larger than a native protein due to the addition of His-tag)
  • Not reactive in: No confirmed exceptions from predicted reactivity are currently known.
  • Nitrogenase is involved in biological fixation of nitrogen to assimilable ammonia.This product is a recombinant protein standard, source: Nostoc/Anabaena 7120.
  • 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)
  • Concentration: after adding 90 µl of milliQ water final concentration of this standard is 0.15 pmoles/ul and this reagent is ready to use and load on a gel.Protein standard buffer composition: Glycerol 10%, Tris Base 141 mM, Tris HCl 106 mM, LDS 2%, EDTA 0.51 mM, SERVA® Blue G250 0.22 mM, Phenol Red 0.175 mM, pH 8.5, 0.1mg/ml PefaBloc protease inhibitor (Roche), 50mM DTT.This standard is ready-to-load and does not require any additions or heating. It needs to be fully thawed and thoroughly mixed prior to using. Avoid vigorous vortexing, as buffers contain detergent. Following mixing, briefly pulse in a microcentrifuge to collect material from cap.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.

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