CNX1/2 | CALNEXIN HOMOLOG 1/2
AS12 2365 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana
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KLH-conjugated synthetic peptide derived from Arabidopsis thaliana CNX1 UniProt: P29402 TAIR: AT5G61790, CNX2 UniProt: Q38798, TAIR: AT5G07340. This peptide is NOT present in calreticulins.
CNX2 60.5/61.4 kD, processing aa 1-25, mature peptides 57.6/58.6 kD
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Reactant: Nicotiana benthamiana
Application: Western Blotting
Pudmed ID: 28394025
Journal: New Phytol
Figure Number: 1B
Published Date: 2017-07-01
First Author: Howden, A. J. M., Stam, R., et al.
Impact Factor: 7.96Open Publication
A simple workflow combining nuclear enrichment with quantitative mass spectrometry allows the study of host nuclear processes during infection. (a) Method overview. Detached leaves from 4?wk?old tomato plants were spray?inoculated with Phytophthora capsici zoospores at a concentration of 500 000 spores ml?1, or water as a control (i). Leaves were harvested 8 and 24 h post?infection and subject to nuclear enrichment (ii). Nuclear protein extracts were generated and fractionated in?gel and digested with trypsin (iii). Peptide samples were subjected to liquid chromatography?tandem mass spectrometry (LC?MSMS) analysis and peptide identification and label?free quantification was carried out using the maxquant and perseus software packages, to identify proteins that were differentially expressed during infection. Nuclear predictions were performed using prediction software on our in?house Galaxy server as described in the Methods. Subsequent data filtering was completed using the r software package (iv and v). Three independent biological replicates were generated. I, infected samples; NI, noninfected samples (v). (b) Successful enrichment of nuclear proteins demonstrated by western blotting with anti?histone H3 antibody and subcellular markers. Protein extracts from enriched nuclear samples were compared to a total protein extract (TP) from tomato leaves. Protein concentrations were adjusted for equal loading. Nonnuclear contamination was assessed by probing with anti?UDP?glucose pyrophosphorylase (UGPase) antibody (cytoplasm) and calnexin homologue 1/2 antibody (endoplasmic reticulum). Samples were also run on gels and stained with Coomassie brilliant blue (CBB) to assess protein loading and Rubisco abundance. The figure shows the results from a single biological replicate. Blots for all three replicates are provided in Supporting Information Fig. S2.
CNX1/2 (calnexin homolog 1/2) is a calcium-binding protein involved in protein folding. It interacts with newly synthesized glycoproteins in the endoplasmic reticulum.
Ekanayake et al. (2021) A. DYNAMIN-RELATED PROTEIN DRP1A functions with DRP2B in plant growth, flg22-immune responses, and endocytosis. Plant Physiol. 2021 Feb 3:kiab024. doi: 10.1093/plphys/kiab024. Epub ahead of print. PMID: 33564884.
Kramer et al. (2020). N6-methyladenosine and RNA secondary structure affect transcript stability and protein abundance during systemic salt stress in Arabidopsis. Plant Direct . 2020 Jul 24;4(7):e00239.doi: 10.1002/pld3.239.
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Butler et al. (2019). Soybean resistance locus Rhg1 confers resistance to multiple cyst nematodes in diverse plant species. Phytopathology. 2019 Aug 12. doi: 10.1094/PHYTO-07-19-0225-R.
Howden et al. (2017), Quantitative analysis of the tomato nuclear proteome during Phytophthora capsici infection unveils regulators of immunity. New Phytol. 2017 Jul;215(1):309-322. doi: 10.1111/nph.14540.
Foley et al. (2017). A Global View of RNA-Protein Interactions Identifies Post-transcriptional Regulators of Root Hair Cell Fate.Dev Cell. 2017 Apr 24;41(2):204-220.e5. doi: 10.1016/j.devcel.2017.03.018.
LaMontagne et al. (2016). Isolation of Microsomal Membrane Proteins from Arabidopsis thaliana. Curr. Protoc. Plant Biol. 1:217-234. doi: 10.1002/cppb.20020.
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