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NBR1 | Autophagy substrate NBR1

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

NBR1 | Autophagy substrate NBR1 in the group Antibodies Plant/Algal  / Environmental Stress / Pathogen attack at Agrisera AB (Antibodies for research) (AS14 2805)
NBR1 | Autophagy substrate NBR1



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Product Information

Immunogen
UBA2 domain of NBR1 of Arabidopsis thaliana, fused with GST, UniProt:Q9SB64, TAIR:AT4G24690
Host Rabbit
Clonality Polyclonal
Purity Serum
Format Lyophilized
Quantity 50 ĩl
Reconstitution For reconstitution add 50 ĩl of sterile water
Storage Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. 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 Immunolocalization (IL), Western blot (WB)
Recommended dilution 1: 1000 (IL), 1 : 500-1 : 5000 (WB)
Expected | apparent MW

75 | 100 kDa

Reactivity

Confirmed reactivity Arabidopsis thaliana, Physcomitrium patens
Predicted reactivity Brassicaceae family

Species of your interest not listed? Contact us
Not reactive in No confirmed exceptions from predicted reactivity are currently known

Application examples

Application examples 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

Reactant: Arabidopsis thaliana (Thale cress)

Application: Western Blotting

Pudmed ID: 29079776

Journal: Nat Commun

Figure Number: 5A

Published Date: 2017-10-27

First Author: Dong, Y., Silbermann, M., et al.

Impact Factor: 13.783

Open Publication

Autophagy is specifically induced by limited S-precursor supply for cysteine biosynthesis. a Autophagy induction in the shoot of serat tko and sir1-1 was determined by immunological detection of the canonical autophagy marker ATG8a and NBR1 with specific antisera. Lipidation of ATG8a (ATG8a-PE) (apparent size: 15–20?kDa) is essential for autophagosome formation and indicated by a significant shift during electrophoresis. NBR1 is a cargo receptor for selective autophagy and consequently degraded in autophagic bodies (D-NBR1) (apparent size: 50–75?kDa). b Level of ATG8a-PE and D-NBR1 shown in a were quantified (n?=?3, mean?ą?s.e.m., one-way ANOVA, *p?

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

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Background

Background
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.

Product citations

Selected references Rodriguez et al. (2020). Autophagy mediates temporary reprogramming and dedifferentiation in plant somatic cells. bioRxiv doi.org/10.1101/747410
Calero-MuÃ?Âąoz et al. (2019). Cadmium induces reactive oxygen species-dependent pexophagy in Arabidopsis leaves. Plant Cell Environ. 2019 Sep;42(9):2696-2714. doi: 10.1111/pce.13597.
Jia et al. (2019). Noncanonical ATG8-ABS3 interaction controls senescence in plants. Nat Plants. 2019 Feb;5(2):212-224. doi: 10.1038/s41477-018-0348-x.
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)

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