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product information
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| background |
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Nitrogenase is involved in biological fixation of nitrogen to assimilable ammonia. Alternative protein names: nitrogenase component II, nitrogenase Fe protein, nitrogenase reductase |
| immunogen |
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KLH-conjugated synthetic peptide derived from known bacterial NifH subunits of bacterial nitrogenase enzymes of the FeMoCo type including Synechoccocus sp. Q2JP78 , Trichodesmium theibautii, Anabaena sp. P33178 and Nostoc sp. Q51296
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| antibody format |
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hen |
polyclonal |
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affinity purified IgY in PBS pH 8.0+ 0.02% sodium azide |
liquid |
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| quantity |
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| storage |
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store at 4°C; make aliquots to avoid working with a stock. Please, remember to spin tubes briefly prior to opening them to avoid any losses that might occur from liquid material adhering to the cap or sides of the tubes. |
| tested applications |
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western blot (WB) |
| related products |
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AS01 021S | NifH protein standard |
| additional information |
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antibody concentration is 1.36 µg/µl recommended secondary antibodies |
application information
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| recommended dilution |
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1: 2 000 (WB) |
| expected | apparent MW |
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27 | 32.5 kDa |
| confirmed reactivity |
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Anabaena PCC7120, Frankia sp. , Nostoc sp, Trichodesmium sp. |
| predicted reactivity |
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Desulfotomaculum reducens (strain MI-1),Clostridium cellobioparum, Frankia sp. (strain EAN1pec),Magnetococcus sp., Methanobacterium thermoautotrophicum, Methanococcus maripaludis, Methylobacterium sp
alpha,gamma,beta proteobacteria, enterobacteria, low GC gram+, high GC gram +, euryachaeotes, Azotobacter vinelandii (Gram-), cyanobacteria |
| not reactive in |
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Synechococcus sp. PCC 7942 and Synechocystis sp. PCC 6803 as NifH protein is not present in those cyanobacterial species |
| additional information |
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This antibody is not suitable for immunolocalization studies on bacterial cultures An enzyme involved in chlorophyll synthesis, present in all cyanobacteria (fixing and non-nitrogen fixing) is a member of the NifH family/superfamily. Agrisera anti-NifH antibody will not show a strong reactivity to this target. |
| selected references |
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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) Küpper et al. (2008). Iron limitation in the marine cyanobacterium Trichodesmium reveals new insights into regulation of photosynthesis and nitrogen fixation. New Phytol. 179: 784-798. |
application example
 Total Trichodesmium sp. protein extract(lanes 6-11, 80 pmol chlorophyll loaded) extracted with PEB (AS08 300), and NifH protein standard (lanes 1-5, 0.05, 0.1, 0.3 0.75 and 1.5 pmol standard loaded) were separated on 4-12% NuPage (Invitrogen) LDS-PAGE and blotted 1h to PVDF. Blots were blocked immediately following transfer in 2% ECL Advance blocking reagent (GE Healthcare) in 20 mM Tris, 137 mM sodium chloride pH 7.6 with 0.1% (v/v) Tween-20 (TBS-T) for 1h at room temperature with agitation. Blots were incubated in the primary antibody at a dilution of 1:40 000 for 1h at room temperature with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed once for 15 min and 3 times for 5 min in TBS-T at room temperature with agitation. Blots were incubated in secondary antibody (anti-hen IgY horse radish peroxidase conjugated, from Abcam) diluted to 1:50 000 in 2% ECL Advance blocking solution for 1h at room temperature with agitation. The blots were washed as above and developed for 5 min with ECL Advance detection reagent
according the manufacturers instructions. Images of the blots were obtained using a CCD imager (FluorSMax, Bio-Rad) and Quantity One software (Bio-Rad).
||| For applications or usage on species others than stated above Agrisera offers a payment-after-testing option. To learn more about this or for any questions on this product, please use the LiveChat option in the left menue bar or contact us at support@agrisera.com
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