NBR1 | Autophagy substrate NBR1

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AS14 2805  |  Clonality: Polyclonal   |  Host: Rabbit   |  Reactivity: Arabidopsis thaliana, Physcomitrella patens


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Item No:
AS14 2805

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product information
NBR1 (Autophagy substrate NBR1) is involved in selective autophagy process of damaged organelles, intracellular microbes, protein aggregates, cellular structures and specific soluble proteins. NBR1 mediates the process as an autophagic adapter. The protein has two UBA domains but only the C-terminal UBA domain bound ubiquitin. Alternative names: At4g24690, Putative uncharacterized protein F22K18.110.
UBA2 domain of NBR1 of Arabidopsis thaliana, fused with GST, UniProt:Q9SB64, TAIR:AT4G24690
Host Rabbit
Clonality Polyclonal
Purity Serum
Format Lyophilized
Quantity 50 µl
Reconstitution For reconstitution 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)
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Additional information
application information
Recommended dilution 1 : 500-1 : 5000 (WB)
Expected | apparent MW

75 | 100 kDa

Confirmed reactivity Arabidopsis thaliana, Physcomitrella patens
Predicted reactivity Brassicaceae family
Not reactive in No confirmed exceptions from predicted reactivity are currently known.
Additional information

Specific extraction method and tissue type needs to be used as described in Minina et al. (2013). Dilution in western blot depends upon amount of NBR1 in the sample.

Selected references Hackenberg et al. (2013). Catalase and NO CATALASE ACTIVITY1 promote autophagy-dependent cell death in Arabidopsis. Plant Cell. 2013 Nov;25(11):4616-26. doi: 10.1105/tpc.113.117192. Epub 2013 Nov 27.
Minina et al. (2013). Autophagy mediates caloric restriction-induced lifespan extension in Arabidopsis. Aging Cell. 2013 Apr;12(2):327-9. doi: 10.1111/acel.12048. Epub 2013 Feb 28. (method description in supplemental materials)
Katsiarimpa et al. (2013). The Deubiquitinating Enzyme AMSH1 and the ESCRT-III Subunit VPS2.1 Are Required for Autophagic Degradation in Arabidopsis.
Svenning et al. (2011). Plant NBR1 is a selective autophagy substrate and a functional hybrid of the mammalian autophagic adapters NBR1 and p62/SQSTM1. Autophagy. 2011 Sep;7(9):993-1010. Epub 2011 Sep 1. (original reference)

Application example

western blot using anti-NBR1 antibodies
Total protein from approximately eight 8-d-old seedlings of Arabidopsis thaliana was extracted with NuPAGE sample buffer (106 mM Tris-HCl pH 8.5, 141 mM Tris Base, 2% lithium dodecyl sulfate, 0.51 mM EDTA pH 8.0, 10% glycerol, 0.22 mM Coomassie Blue G250, 0.166 mM Phenol Red) with 50 mM dithiothreitol and denatured at 100°C for 5 min. Proteins were separated by electrophoresis on a Bolt 10% Bis-Tris Plus gel (Invitrogen) and transferred for 40 min at 24 V to an Amersham Protran 0.45 µm nitrocellulose membrane (GE Healthcare Life Sciences) using semidry transfer. The membrane was blocked with 8% milk in TBS-T for 2 h at 4°C with agitation. The membrane was incubated in the primary antibody at a dilution of 1:4000 overnight at 4°C with agitation in 8% milk in TBS-T. The antibody solution was decanted and the blot was rinsed briefly twice, then washed 3 times for 5 min in 8% milk in TBS-T at 4°C with agitation. The membrane was incubated in secondary antibody (horse radish peroxidase conjugated goat anti-rabbit IgG) diluted to 1:5000 in 8% milk in TBS-T for 4 h at 4°C with agitation. The membrane was washed 3 times for 5 min in TBS-T at 4°C with agitation and incubated for 2 min with WesternBright chemiluminescent detection reagent (Advansta). Exposure time was 30 seconds.

Courtesy of Pierce Young, Rice University, USA

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