PEC1 | Plastid Envelope Channel 1
AS21 4528 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana
compartment marker of chloroplast envelope membrane
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Product Information
Immunogen
The soluble domain 466 (M244 until stop codon, ≈ 64 kDa) was cloned into pET16b and transformed into BLR 21 for 467 expression in Escherichia coli UniProt: Q8VZM7-1, TAIR: At5g02940
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
Western blot (WB)
Recommended dilution
1 : 1000 (WB)
Expected | apparent MW
100 | 75 kDa
Reactivity
Confirmed reactivity
Arabidopsis thaliana
Predicted reactivity
Camelina sativa, Capsella rubella, Nicotiana benthamiana, Noccaea caerulescens, Pisum sativum, Raphanus sativus,
Species of your interest not listed? Contact us
Species of your interest not listed? Contact us
Application examples
Application examples
Samples:
1 – 2,5 ug of Arabidopsis thaliana mutant pec1-1pec2-1
2 – 2,5 ug of Arabidopsis thaliana mutant pec1-2pec2-2
3 – 2,5 ug of Arabidopsis thaliana whole leaf extract
Arabidopsis thaliana tissue was frozen with liquid nitrogen and ground to a fine powder with mortar and pestle. Protein was extracted by mixing with extraction buffer (200 mM tris pH 8.0, 4% (w/v) sodium dodecyl sulfate) to 0.5 g fresh weight/mL and heating at 80°C for 8 min. 5 µl protein extract were separated on 10 % SDS-PAGE and blotted at 75v for 45min to nitrocellulose (pore size of 20 um), using wet transfer. Blot was blocked with 5% milk for 1h/RT with agitation. Blot was incubated in the primary antibody at a dilution of 1: 1000 ON/4°C in 5% Milk-TBS-T with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed once for 15 min and 3 times for 5 min in TBS-T at RT with agitation. Blot was incubated in matching secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1:10 000 in for 1h/RT with agitation. The blot was washed as above and developed for 1 min with chemiluminescent detection reagent following manufacture's recommendations. Exposure time was 120 seconds.
Courtesy of Carsten Voelkner, Kunz Lab
Samples:
1 – 2,5 ug of Arabidopsis thaliana mutant pec1-1pec2-1
2 – 2,5 ug of Arabidopsis thaliana mutant pec1-2pec2-2
3 – 2,5 ug of Arabidopsis thaliana whole leaf extract
Arabidopsis thaliana tissue was frozen with liquid nitrogen and ground to a fine powder with mortar and pestle. Protein was extracted by mixing with extraction buffer (200 mM tris pH 8.0, 4% (w/v) sodium dodecyl sulfate) to 0.5 g fresh weight/mL and heating at 80°C for 8 min. 5 µl protein extract were separated on 10 % SDS-PAGE and blotted at 75v for 45min to nitrocellulose (pore size of 20 um), using wet transfer. Blot was blocked with 5% milk for 1h/RT with agitation. Blot was incubated in the primary antibody at a dilution of 1: 1000 ON/4°C in 5% Milk-TBS-T with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed once for 15 min and 3 times for 5 min in TBS-T at RT with agitation. Blot was incubated in matching secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1:10 000 in for 1h/RT with agitation. The blot was washed as above and developed for 1 min with chemiluminescent detection reagent following manufacture's recommendations. Exposure time was 120 seconds.
Courtesy of Carsten Voelkner, Kunz Lab
Additional information
Additional information
This antibody is recognizing PEC1, but not PEC2 or DMI1
Background
Background
The two Arabidopsis POLLUX-like homologs PEC1 and PEC2 represent plastid envelope membrane cation channels with K + conductivity that are required for the stress triggered Ca 2+ release into the stroma.
Product citations
Selected references
Völkner et al (2021) Two plastid POLLUX ion channel-like proteins are required for stress-triggered stromal Ca2+release, Plant Physiology, Volume 187, Issue 4, December 2021, Pages 2110–2125, https://doi.org/10.1093/plphys/kiab424
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