NifH | Nitrogenase iron protein
AS01 021A | Clonality: Polyclonal | Host: Hen | reactivity: [global antibody] for bacteria
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|Recommended dilution||1 : 500 (IHC), 6 ĩg/ml (IF), 1 : 2000 (WB)|
|Expected | apparent MW||
27 | 32.5 kDa
|Confirmed reactivity||Anabaena PCC7120, Clostridium butyricum, Codakia orbicularis, Cylindrospermopsis raciborskii CS-505, Nostoc sp, Rhodopseudomonas palustris, Trichodesmium sp., nodules of Trifolium repens L.|
Azotobacter vinelandii (Gram-), Bradyrhizobium japonicum, Cyanobacteria, Desulfotomaculum reducens (strain MI-1),Clostridium cellobioparum, Enterobacter genera, euryachaeotes, Klebsiella pneumonia, Magnetococcus sp., Methanobacterium thermoautotrophicum, Methanococcus maripaludis, Methylobacterium sp., Mesoorhizobium loti, Rhodopseudomonas palustris TIE-1 strain, alpha,gamma,beta proteobacteria, enterobacteria, low GC gram+, high GC gram +, able to fix atmoshperic nitrogen, Rhizobium meliloti
|Not reactive in||
Synechococcus sp. PCC 7942 and Synechocystis sp. PCC 6803 as NifH protein is not present in those cyanobacterial species, Frankia sp.
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||König et al. (2016). Nitrogen fixation in a chemoautotrophic lucinid symbiosis. Nat Microbiol. 2016 Oct 24;2:16193. doi: 10.1038/nmicrobiol.2016.193.
Liberti et al. (2015). Bacterial symbiont sharing in Megalomyrmex social parasites and their fungus-growing ant hosts. Mol Ecol. 2015 Apr 24. doi: 10.1111/mec.13216.
Calusinska et al. (2015). Genome-wide transcriptional analysis suggests hydrogenase- and nitrogenase-mediated hydrogen production in Clostridium butyricum CWBI 1009. Biotechnology for Biofuels (2015) 8:27.
Moirangthem et al. (2014). A high constitutive catalase activity confers resistance to methyl viologen-promoted oxidative stress in a mutant of the cyanobacterium Nostoc punctiforme ATCC 29133. Appl Microbiol Biotechnol. 2014 Jan 3.
Chen et al. (2013). Improving conversion efficiency of solar energy to electricity in cyanobacterial PEMFC by high levels of photo-H2 production. Int J of Hydrogen Energy (2013):1-8.(Anabaena, western blot)
Plominsky et al. (2013). Dinitrogen Fixation Is Restricted to the Terminal Heterocysts in the Invasive Cyanobacterium Cylindrospermopsis raciborskii CS-505. PLOS ONE, Open Access.
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) 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).
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