NifH | nitrogenase iron protein

345 €

AS01 021A | clonality: polyclonal | host: hen | reactivity: [global antibody] for bacteria

PRODUCT INFORMATION IN PDF

product information

background

Nitrogenase is involved in biological fixation of atmospheric nitrogen to ammonia. Alternative protein names: nitrogenase component II, nitrogenase Fe protein, nitrogenase reductase, FeMoCo-nitrogenase.

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

antibody format

hen

polyclonal

affinity purified IgY in PBS pH 8.0+ 0.02% sodium azide

liquid

quantity

200 µg

storage

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

western blot (WB)

related products

AS01 021S | NifH protein standard

additional information

antibody concentration is 1.36 µg/µl

recommended secondary antibodies

application information
recommended dilution		1: 2 000 (WB)
expected \| apparent MW		27 \| 32.5 kDa
confirmed reactivity		Anabaena PCC7120, Cylindrospermopsis raciborskii CS-505, Nostoc sp, Rhodopseudomonas palustris, Trichodesmium sp., nodules of Trifolium repens L.
predicted reactivity		Desulfotomaculum reducens (strain MI-1),Clostridium cellobioparum, Frankia sp. (strain EAN1pec), Magnetococcus sp., Methanobacterium thermoautotrophicum, Methanococcus maripaludis, Methylobacterium sp , Rhodopseudomonas palustris TIE-1 strain, alpha,gamma,beta proteobacteria, enterobacteria, low GC gram+, high GC gram +, euryachaeotes, Azotobacter vinelandii (Gram-), Klebsiella pneumonia, cyanobacteria, Enterobacter genera, able to fix atmoshperic nitrogen
not reactive in		Synechococcus sp. PCC 7942 and Synechocystis sp. PCC 6803 as NifH protein is not present in those cyanobacterial species
additional information		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		Plominsky et al. (2013). Dinitrogen Fixation Is Restricted to the Terminal Heterocysts in the Invasive Cyanobacterium Cylindrospermopsis raciborskii CS-505. PLOS ONE, Open Access. Richier et al. (2012). Abundances of Iron-Binding Photosynthetic and Nitrogen-Fixing Proteins of Trichodesmium Both in Culture and In Situ from the North Atlantic. Plos one. 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)

application example

western blot using anti-NifH hen antibody

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).

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