CURT1A | Curvature thylakoid 1A

AS08 316 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana, Pisum sativum

Product Information

Immunogen

KLH-conjugated synthetic peptide derived from Arabidopsis thaliana CURT1A sequence, TAIR: AT4G01150

Host Rabbit

Clonality Polyclonal

Purity Affinity purified serum in PBS, pH 7.4

Format Lyophilized

Quantity 50 µg

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 Blue Native PAGE (2D BN-PAGE), Western blot (WB)

Recommended dilution 1 : 1000 (WB)

Expected | apparent MW

17.6 | 11 kDa (due to N-terminal processing)

Reactivity

Confirmed reactivity Arabidopsis thaliana, Pisum sativum

Predicted reactivity Nicotiana tabacum, Zea mays

Not reactive in

Hordeum vulgare, Phaseolus coccineus

Application examples

Application example

The dillution series of Hordeum vulgare (4; 2; 1 µg Chl) and Arabidopsis thaliana (4; 2; 1 µg Chl) thylakois were separated on 12% Criterion XT Bis-Tris SDS-PAGE (BioRad) gels and blotted for 25min 100V to PVDF membrane. Blot was blocked with 5% fat free skimmed milk in PBS-T for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1:1000 overnight with agitation in 4˚C. The antibody solution was decanted and the blot was washed 3 times for 5 min in PBS-T at RT with agitation. Blot was incubated in secondary antibody (swine anti-rabbit IgG horse radish peroxidase conjugated, from Dako) diluted to 1: 5000 in 1% fat free skimmed milk in PBS-T for 1h at RT with agitation. The blot was washed 5 min in PBS-T and 1 min in PBS developed for 5 min with ECL according to the manufacturers instructions. Exposure time was 60 min.

Courtesy of Dr. Marta Powikrowska, University of Copenhagen, Danmark

Western blot using anti-CURT1A antibodies

Plant species: Arabidopsis LD – Arabidopsis thaliana grown under long day conditions Arabidopsis SD – Arabidopsis thaliana grown under short day conditions Runner bean – Phaseolus coccineus L. Pea – Pisum sativum L. Experimental procedure: Samples of isolated thylakoids containing 50 µg of chlorophyll were denatured with 150 mL double diluted Roti®-Load 1 (ROTH, Art.-Nr. K929.1) at 95 °C for 2 min. Denatured samples containing 0.5 and 1 µg of chlorophyll were loaded in the gel wells and separated on 14% SDS-PAGE gels and blotted for 45 min at 105 V to PVDF membrane using wet transfer. Blot was blocked with 5% Amersham™ ECL Prime Blocking Agent (cat no: RPN418) in TBS-T for 30 min at room temperature (RT) with agitation. The blot was incubated with the primary antibody at a dilution of 1:1000 overnight at 4˚C with agitation. The antibody solution was decanted and the blot was washed 2 times for 5 min in TBS-T at RT with agitation. The blot was incubated using a secondary antibody (goat anti-rabbit IgG HRP conjugated, from Agrisera, AS09 602) diluted to 1: 25000 in 1% Blocking Agent in TBS-T for 1h at RT with agitation. The blot was washed 2 times for 5 min in TBS-T and developed for 5 min with chemiluminescent reagent, according to instructions. Exposure time was 12 minutes in C-Digit chemiluminescence scanner (Li-COR).

Dr Radosław Mazur, Faculty of Biology, University of Warsaw, Poland

Additional information

Related products

Plant protein extraction buffer

Background

Curvature thylakoid 1A (CURT1A) belongs to a protein family, conserved in plants and cyanobaceria. There are four Arabidopsis thaliana CURT1 proteins: CURT1A,B,C and D. It is proposed that CURT1 proteins modify thylakoid architecture by inducing membrane curvature at grana margins.

Product citations

Selected references Armbruster et al. (2013). Arabidopsis CURVATURE THYLAKOID1 Proteins Modify Thylakoid Architecture by Inducing Membrane Curvature. 2013 Jul;25(7):2661-78. doi: 10.1105/tpc.113.113118. Epub 2013 Jul 9.