Nitrate Reductase, Assimilatory (NR) antibody

Antigen: Nitrate Reductase, Assimilatory (NR)

Clonality: Polyclonal

Host: Rabbit

Reactivity: Arabidopsis thaliana, Barley (Hordeum vulgare), Red algae (Gracilaria), Thalassiosira

Application: Western Blotting (WB)

Catalog no.: ABIN334562

Additional Information

Immunogen: KLH-conjugated synthetic peptide derived from conserved domain in NADH-NRprotein sequences including A.thaliana NR1 P11832 and NR2 P11035

Format: Lyophilized

Description: Assimilatory nitrate reductase (NR, EC.1.6.6.1) catalyses the reduction of nitrateto nitrite in the cytoplasm. Plants contain 2 forms of NR: NADH-NR (mostcommon form in plants and algae, predominantly found in green tissues) andNAD(P)H-NR (uses NADH or NADPH as the electron donor, constitutivelyexpressed in plants at a low level). NADH-NR is a homodimer of two identicalsubunits (100-115 kDa each, hold together by a Mo-cofactor) each of them codedby up to three genes (NR1-3, NIA1-NIA3).

Specificity: Predicted reactivity: dicots including: Glycine max, Lycopersicum esculentum, Nicotiana tabacum,Ricinus communis, Solanum tuberosum, monocots including: Oryza sativa, Zeamays, moss: Physcomitrella patens, algae Chlorella vulgaris, Dunaliella salina,marine diatoms.

Application Details

Application Notes: Recommended Dilution 1: 1000 (WB).

Purity: Affinity purified serum

Storage: store lyophilized/reconstituted at -20°C, once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please, remember to spin tubes briefly prior toopening them to avoid any losses that might occur from lyophilized materialadhering to the cap or sides of the tubes.

Research Area: Plant/Algal Physiology

Restrictions: For Research Use only

Images

Detection of NR in protein extracts from leaves of Columbia (1) and NR knockout(2-3) mutants of Arabidopsis thaliana. Each lane was loaded with the protein contained in2mg fresh weight leaf tissue (~15ug total protein). Mutants contained 56% (2) and 5% (3)of the NR activity of Columbia. Proteins were extracted in 0.1M Hepes pH 7.5, 1mM EDTA,0.1% Triton X-100, 2 mM dithiothreitol, 0.7% Protease cocktail mix for plants (Sigma), andseparated on 12% SDS-PAGE and blotted for 1h to nitrocellulose. Filters were blocked for1h with 2% BSA in TBS-T (0.1% TWEEN 20) and probed with anti-NR antibodies AS08310 in dilution 1:1000, 1h) incubation time and secondary anti-rabbit (1: 4 000, 1 h)antibody (HRP conjugated, DAKO) in TBS-T containing 2% BSA. Antibody incubationswere followed by three washings in TBS-T (15 min each). All steps were performed at RTwith agitation. Signal was detected on blue light sensitive film (exposure time 1 min) withstandard ECL (GE Healthcare). The image was captured with GS800 calibrateddensitometer (BioRad, UK Ltd). Black arrow shows gel origin, blue arrows show bands ofcross reactivity. Courtesy Dr. Alison Kingston-Smith and Teri Davies, Aberystwyth University 1

anti-Nitrate Reductase, Assimilatory (NR) antibody (Image 2)

anti-Nitrate Reductase, Assimilatory (NR) antibody (Image 3)