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Pex14p | Peroxisomal marker

265 €
Buy 2 items of this product for 198 €/each
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AS08 372 | clonality: polyclonal | host: rabbit | reactivity: Arabidopsis thaliana, Magnaporthe oryzae

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Item No:
AS08 372

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product information
Background

Pex14p has a protein transporter activity and is involved in protein targeting into peroxisome. Submitted protein name: Genomic DNA, chromosome 5, P1 clone:MQB2

Immunogen Recombinant, soluble N-terminal domain of Arabidopsis thaliana Pex14p (Q9FE40, At5g62810) that mediates PEX5 and PEX19 binding. The transmembrane and coiled-coil domains of PEX14 were replaced with a dual StrepII-His6 tag.
Host Rabbit
Clonality Polyclonal
Clone
Purity Affinity purified serum
Format Lyophilized
Quantity 50 µg
Reconstitution For reconstitution add 25 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)
Related products

AS10 711 | DEG15 | endopeptidase, peroxisomal marker - with wider species reactivity than Arabidopsis thaliana

AS11 1797 | HPR | hydroxypyruvate reductase (peroxysomal matrix marker)

Plant protein extraction buffer

Secondary antibodies

Additional information Pex14p is prone to degradation.
application information
Recommended dilution 1: 10 000 with standard ECL (WB)
Expected | apparent MW

55.5 | 75-65 kDa (depending on the gel system)

Confirmed reactivity Arabidopsis thaliana
Predicted reactivity Arabidopsis thaliana, Magnaporthe oryzae
Not reactive in no confirmed exceptions from predicted reactivity known in the moment
Additional information Antibodies will detect Pex14 protein in both light- and dark-grown seedlings grown on petri dishes and in rosette leaves of adult plants grown in soil.
Selected references Zhong et al. (2016). MoDnm1 Dynamin Mediating Peroxisomal and Mitochondrial Fission in Complex with MoFis1 and MoMdv1 Is Important for Development of Functional Appressorium in Magnaporthe oryzae. PLoS Pathog. 2016 Aug 24;12(8):e1005823. doi: 10.1371/journal.ppat.1005823. eCollection 2016.
Li et al. (2015). A Chaperone Function of NO CATALASE ACTIVITY1 Is Required to Maintain Catalase Activity and for Multiple Stress Responses in Arabidopsis. Plant Cell. 2015 Feb 19. pii: tpc.114.135095.
Woodward et al. (2014). A viable Arabidopsis pex13 missense allele confers severe peroxisomal defects and decreases PEX5 association with peroxisomes. Plant Mol Biol. 2014 Jul 10.
McMichael et al. (2013). Mediation of Clathrin-Dependent Trafficking during Cytokinesis and Cell Expansion by Arabidopsis STOMATAL CYTOKINESIS DEFECTIVE Proteins. Plant Cell, Oct. 31.
Farmer et al. (2013).Disrupting Autophagy Restores Peroxisome Function to an Arabidopsis lon2 Mutant and Reveals a Role for the LON2 Protease in Peroxisomal Matrix Protein Degradation. Plant Cell, Oct 31.

application example

western blot with plant peroxisomal marker antibody

Twenty 5-day-old light grown Arabidopsis thaliana seedlings (wild type, the pex14-2 null allele, and the pex14-1 truncation allele) were ground with a pestle in a 1.5 mL tube on dry ice following the addition of a double volume (60 μL) of NuPAGE 2x loading buffer (Invitrogen). After centrifugation, 20 μL of supernatant was trasnferred to a fresh tube with 2.1 μL 0.5 M DTT and boiled at 100 °C for 5 mins. Samples were loaded onto NuPAGE 10% Bis-Tris gel (Invitrogen) next to Cruz Marker (Santa Cruz Biotechnology). After electrophoresis, proteins were transferred for 30 mins at 24 V to a Hybond ECL nitrocellulose membrane (Anersham Pharmacia Biotech) using NuPAGE transfer buffer (Invitrogen). The blot was blocked for 1 h 4 °C in 8% non-fat dry milk in TBS-T (blocking buffer),and incubated overnight wiht agitation at 4 °C with primary antibody (1:10 000 rabbit anti-PEX14, AS08 372, Agrisera) diluted in blocking buffer. The antibody soliution was decanted and the blot was rinsed 3 times, 5 min each, with blocking buffer at 4 °C. The blot was incubated with horseradish peroxidase-conjugated anti-rabbit secondary antibody (1:5000, Santa Cruz Biotechnology), diluted in blocking buffer for 4 h at 4 °C with agitation. The blot was washed with 3 times, 5 min each, with TBS-T and developed with Western Bright reagent (Advanta), 1:10 dilution in water.

Courtesy Sarah Burkhart and Bonnie Bartel, Rice University, USA.


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