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AGO set
Tag Antibody Set 

LOX | Lipoxygenase (Affinity purified)

AS06 128A  |  Clonality: Polyclonal  |  Host: Rabbit  |  Reactivity: A.thaliana, G. max, S. tuberosum

LOX | Lipoxygenase (Affinity purified) in the group Antibodies for Plant/Algal  / Developmental Biology / Lipid metabolism at Agrisera AB (Antibodies for research) (AS06 128A)

PRODUCT INFORMATION IN PDF

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Datasheet Product citations Protocols Add review

Product Information

Immunogen

native lipoxygenase, type I-B, purified from Glycine max (Sigma, product number L7395) UniProt: P08170

Host Rabbit
Clonality Polyclonal
Purity Affinity purified serum in PBS, pH 7.4
Format Lyophilized
Quantity 50 µg
Reconstitution For reconstitution of lyophilized unit please add 50 µl of sterile water.
Storage Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please, remember to spin tubes briefly prior to opening them to avoid any losses that might occur from lyophilized material adhering to the cap or sides of the tubes.
Tested applications Western blot (WB)
Recommended dilution 1 : 5000 (WB)
Expected | apparent MW

54 (subunit), 108 (native enzyme)

Reactivity

Confirmed reactivity Arabidopsis thaliana, Glycine max, Solanum tuberosum
Predicted reactivity

Cucumis sativus, Lathyrus undulatus, Malus x domestica

Not reactive in No confirmed exceptions from predicted reactivity are currently known.

Application examples

Application examples Application example

western blot using anti-LOX antibodies

10 µg of total protein from Arabidopsis thaliana (1), native LOX protein from Glycine max (2,3) total protein from all the samples were extracted with Protein Extraction Buffer PEB (AS08 300). Samples were diluted with 1X sample buffer (NuPAGE LDS sample buffer (Invitrogen) supplemented with 50 mM DTT and heat at 70°C for 5 min and keep on ice before loading. Protein samples were separated on NuPAGE 4-12% Tris-Bis gel (Invitrogen) LDS-PAGE and blotted for 1h to 1.5h on PVDF using tank transfer. Blots were blocked immediately following transfer in 2% blocking reagent or 5% non-fat milk dissolved 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: 5 000 (in blocking reagent) for 1h at room temperature with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, and then washed 1x15 min and 3x5 min with TBS-T at room temperature with agitation. Blots were incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, recommended secondary antibody AS09 602) diluted to 1:25 000 in blocking reagent for 1h at room temperature with agitation. The blots were washed as above. The blot was developed for 5 min with chemiluminescent 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).



Western blot using anti-LOX antibodies on potato tuber flesh

Proteins were extracted from tuber flesh of Solanum tuberosum cv. Russet Burbank with 0.1 M Tris HCl (pH=8.0), 5% sucrose (m/v), 2% (m/v) SDS, protease inhibitors (PMSF 1mM). Samples were heated 95°C 5 min, and 10 μg of total protein was resolved in 12% SDS PAGE and blotted to PVDF membrane for 1h-1.5h using tank transfer. Blots were blocked with a skimmed milk 4% (m/v) in T-TBS (1.5h) at RT with agitation. Primary antibodies (AS06 128A) were applied overnight +4°C in dilution 1:5000 with agitation. After washing with T-TBS 2 times, membrane was incubated with secondary antibodies (Goat Anti-Rabbit HRP conjugated) 1:10 000 for 1 hour at RT. Blot was washed as above and developed with chemiluminescence detection reagent, for 5 – 10 minutes. Exposure time – 7.000 seconds.

Courtesy of Msc. Iauhenia Isayenka, Laboratory of Nathalie Beaudoin, University of Sherbrooke, Canada

Additional information

Related products

Related products

AS06 128 | Anti-LOX | lipoxygenase, rabbit antibodies (not purified serum)

AS07 258 | Anti-LOX-C | chloroplastic lipoxygenase, rabbit antibodies

AS08 340 | Anti-LOX2 | lipoxygenase 2 protein, rabbit antibodies

Plant protein extraction buffer

Secondary antibodies

Background

Background

Lipoxygenases (LOXs; EC 1.13.11.12, synonym: lipoxydases) are a family of enzymes that catalyze oxygenation of polyunsaturated fatty acids (PUFAs) into lipidhydroperoxides (LOOHs) involved in responses to stresses. LOXs has been found to play a role in plant growth and development, senescence as well as can be activated in response to environamental stress (drought, heavy metals).Synonymes: linoleate, oxygen oxidoreductase.

Product citations

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