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{{/_source.additionalInfo}}Anti-NifH | Nitrogenase iron protein
(Cat#: AS01 021A)
Description
- Immunogen: 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
- Host: Chicken
- Clonality: Polyclonal
- Purity: Immunogen affinity purified IgY in PBS pH 8 and 0.02 % sodium azide.
- Format: Liquid at 1.28 mg/ml
- Quantity: 100 µg
- Storage: Store at 4°C; make aliquots to avoid working with a stock. Please remember to spin the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.
- Tested applications: Immunofluorescence (IF), Immunohistochemistry (IHC), Western blot (WB)
- Recommended dilutions: 1 : 500 (IHC), 6 µg/ml (IF), 1 : 2000 (WB)
- Expected | apparent MW: 27 | 32.5 kDa
- Confirmed reactivity: Anabaena PCC7120, Clostridium butyricum, Codakia orbicularis, Cylindrospermopsis raciborskii CS-505, Dolichospermum sp., Nostoc sp, Rhodopseudomonas palustris, Trichodesmium sp., nodules of Trifolium repens L., Vibrio natriegens ATCC 14048
- Not reactive in: Synechococcus sp. PCC 7942 and Synechocystis sp. PCC 6803 as NifH protein is not present in those cyanobacterial species, Frankia sp.
- Nitrogenase is involved in biological fixation of atmospheric nitrogen to ammonia. Alternative protein names: nitrogenase component II, nitrogenase Fe protein, nitrogenase reductase, FeMoCo-nitrogenase.
- Zhou et al. (2024). Inorganic nitrogen inhibits symbiotic nitrogen fixation through blocking NRAMP2- mediated iron delivery in soybean nodules. Nat Commun . 2024 Oct 17;15(1):8946. doi: 10.1038/s41467-024-53325-y. Wen et al. (2024). Effects of CO2 on the nitrogen isotopic composition of marine diazotrophic cyanobacteria. Geophysical Research Letters.Ito et al. (2024). Improvement of the nitrogenase activity in Escherichia coli that expresses the nitrogen fixation-related genes from Azotobacter vinelandii. Biochem Biophys Res Commun. 2024 Jul 3:728:150345.doi: 10.1016/j.bbrc.2024.150345.Fernández-Juárez et al. (2023), Biofilm formation and cell plasticity drive diazotrophy in an anoxygenic phototrophic bacterium. Appl Environ Microbiol. 2023 Nov ;89(11):e0102723. doi: 10.1128/aem.01027-23.Santana-Sanchez, et al. (2023) Flv3A facilitates O2 photoreduction and affects H2 photoproduction independently of Flv1A in diazotrophic Anabaena filaments. New Phytol. 2023;237(1):126-139. doi:10.1111/nph.18506Chen et al. (2022) Exogenous hydrogen sulphide alleviates nodule senescence in Glycine max-Sinorhizobium fredii symbiotic system, Preprint from Research Square, 22 Jul 2022, DOI: 10.21203/rs.3.rs-1752770/v1Li et al. (2022), The effects of Ni availability on H2 production and N2 fixation in a model unicellular diazotroph: The expression of hydrogenase and nitrogenase. Limnol Oceanogr, 67: 1566-1576. https://doi.org/10.1002/lno.12151He et al. (2021) Vegetative cells may perform nitrogen fixation function under nitrogen deprivation in Anabaena sp. strain PCC 7120 based on genome-wide differential expression analysis. PLoS One. 2021 Mar 4;16(3):e0248155. doi: 10.1371/journal.pone.0248155. PMID: 33662009; PMCID: PMC7932525. (Immunolocalization)Liu et al. (2020). A VIT-like transporter facilitates iron transport into nodule symbiosomes for nitrogen fixation in soybean. New Phytol . 2020 Mar 2. doi: 10.1111/nph.16506.
- 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.In photobionts like Anabaena sp., low nitrate growth is required to turn on the NifH expression to high enough levels to detect NifH protein. Immunofluorescence protocol Insect dissected tissues (digestive tract, fat body, carrying NifH positive bacteria) of large workers were fixed in cold methanol (20 min, -20°C) and then permeabilized in cold acetone (5 min, -20°C). Samples were subsequently rinsed three times with PBS with 0.1 % Triton-X 100 at RT (PBST) and incubated for 5 minutes in PBST. This was followed by incubation of tissues for 1 hr with 6 ug/ml affinity purified anti-NifH antibody (Agrisera, AS01 021A) diluted in PBS-TBSA (PBS, 0.1 % v/v Triton-X-100, 1 mg/ml BSA) and 3 washings with PBST. Samples were then incubated in the dark with a goat anti-chicken IgY conjugated to Dylight 488 (Pierce, SA5-10070) for 45 min and were washed twice (PBS, 0.1%v/v Triton-X-100). Finally, the tissues were mounted in Vectashield medium containing DAPI (Vector Laboratories, H-1500) and viewed under a SP5 Leica confocal microscope with 10X and 63X objectives. Courtesy of Drs. Panagiotis Sapountzis and Mariya Zhukova, University of Copenhagen, Danmark
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