G4 | Chlorophyll synthase (chloroplastic)

AS14 2793 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Hordeum vulgare

Product Information

Immunogen

KLH-conjugated peptide derived from Arabidopsis thaliana chlorophyll synthase, UniProt: Q38833, TAIR: AT3G51820

Host Rabbit

Clonality Polyclonal

Purity Serum

Format Lyophilized

Quantity 50 µl

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 Western blot (WB)

Recommended dilution 1 : 1000 (WB)

Expected | apparent MW

41.8 kDa (further processed into a mature form)

Reactivity

Confirmed reactivity Hordeum vulgare

Predicted reactivity Arabidopsis thaliana, Avena sativa, Camelia sinensis, Cocomyxa subellipsoidea, Gossypium hirsutum, Micromonas pusilla, Nicotiana tabacum, Oryza sativa, Populus trichocarpa, Ricinus communis, Triticum urartu, cyanobacteria

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

Barley seeds were grown for 4.5 days in the dark before they were harvested and intact plastids were isolated in light according to Eichacker et al. 1996. The concentration of plastids was determined by counting in a Thoma counting chamber. The membrane complexes were isolated according to Reisinger et al. 2008 before 3x SB buffer (2% (w/v) SDS, 10% (w/v) sucrose, 0.03% (w/v) bromphenol blue, 66 mM NaCO3, 66 mM dithiothreitol) was added. The proteins were denatured for 2 min at 72 °C. The membrane fractions were loaded onto a 12.5 % SDS-acrylamide gel, giving approximately 2.5x107 plastids per lane alongside with Magic Marc (LC5602, Life technologies). The gel was then run at 175 V with 1xSDS running buffer. The proteins were then transferred onto a nitrocellulose membrane by using semi-wet transfer and Towbin buffer w/10 % methanol. The proteins were transferred at 2 mA/cm2 for 1 hour. The membrane was blocked with TBS-milk, washed 3x5 min in TBS-tween and the primary antibody (Chlorophyll synthase, AS14 2793 from Agrisera) was added at a concentration of 1:1000. The membrane was incubated ON at 4 °C (approximately 18 h) before washing with TBS-tween and incubation for 1h at RT with the secondary antibody diluted 1:12 500 (Agrisera AS09 602). After washing, the membrane was incubated in an ECL1/ECL2 solution (100 mM Tris, 2.5 mM Luminol, 0.4 mM pCoumaracid/100 mM Tris, 0.0183 % Hydrogenperoxide) before exposure to a film for 7 min. The film was then developed and fixed using chemicals from Kodak.

Courtesy of Eli Drange Vee, Staff Engineer, Centre for Organelle Research, University of Stavanger, Norway

Additional information

Related products

AS08 300 | protein extraction buffer for plant and algal samples

Plant protein extraction buffer

Background

G4 (Chlorophyll synthase) is an enzyme which performs estrification of chlorophyllide (a and b), which is the last step of chlorophyll biosynthesis. Alternative names: polyprenyl transferase, protein G4.