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Available ordering format
Lyophilized
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Immunogen
to be determined
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Raised in
N/A
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Clonality
N/A
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Clone
N/A
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Purification
Affinity purified
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How to reconstitute
For reconstitution add 225 µl of milliQ water
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Storage condition
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.
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Verified applications
western blot (WB)
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Connected products
AS01 021A | matching antibody: anti-NifH hen global antibodiescollection of other protein standardscollection of other global antibodiescollection of antibodies to proteins involved in nitrogen metabolismAlgal protein extraction buffer
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Recommended dilutions for use
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.
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Molecular weight expected аpparent
34 kDa (larger than a native protein due to the addition of His-tag)
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Verified reactivity
to be determined
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Possible reactivity
to be determined
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No reactivity
no confirmed exceptions from predicted reactivity known in the moment
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Supplementary information
Concentration: after adding 225 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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References
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)
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Scientific context
Nitrogenase is involved in biological fixation of nitrogen to assimilable ammonia.This product is a recombinant protein standard, source: Nostoc/Anabaena 7120.
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Notes
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
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Protein number
Refer to NCBI
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TAIR number
Refer to NCBI
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Description
Isotype or positive controls by peptides, antibodies and deactivated samples. Positive controls are the same as the target vector or antibody or protein and can be spiked to the sample before the analysis starts.
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Group
positif
