Lhca4 | PSI type IV chlorophyll a/b-binding protein

BSA-conjugated synthetic peptide derived from the Lhca4 protein ofArabidopsis thaliana UniProt: P27521, TAIR: At3g47470. This sequence is highly conserved in Lhca4 proteins of angiosperms (monocots and dicots) and gymnosperms.

27.7 | 21 kDa for Arabidopsis thaliana

1 µg of chlorophyll from Pisum sativum (1), Mesembryanthemum crystallinum (2), mesophyll (3) and bundle sheath (4) of Zea mays, mesophyll (5) and bundle sheath (6) of Echinochloa crus-galli chloroplasts extracted with 0.4 M sorbitol, 50 mM Hepes NaOH, pH 7.8, 10 mM NaCl, 5 mM MgCl2 and 2 mM EDTA were loaded to lanes. Samples were denatured with Laemmli buffer at 75 0C for 5 min and were separated on 12% SDS-PAGE, and blotted 30 min to PVDF using wet transfer. Blot was blocked with 5% milk for 2h at room temperature (RT) with agitation. Blot was incubated in the primary antibody Anti-Lhca4 (LOT 1908) at a dilution of 1: 3000 in 1% milk in TBS-T overnight at 4°C with agitation. The antibody solution was decanted and the blot was washed 4 times for 5 min in TBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG HRP conjugated, from Agrisera, AS09 602, LOT 1905) diluted to 1:20 000 in 1 % milk in TBS-T for 1h at RT with agitation. The blot was washed 5 times for 5 min in TBS-T and 2 times for 5 min in TBS, and developed for 1 min with 1.25 mM luminol, 0.198 mM coumaric acid and 0.009% H₂O₂ in 0.1 M Tris- HCl, pH 8.5. Exposure time in ChemiDoc System was 30 seconds.

Courtesy of Dr. Wioleta Wasilewska-Dębowska, Warsaw University, Poland

AS01 011 | A set of 10 plant anti-Lhca and anti-Lhcb antibodies

AS01 011 Chlamydomonas | A set of anti-Lhc antibodies for Chlamydmonas

Available antibodies against pigment-binding proteins- LHC

recommended secondary antibody

Plant protein extraction buffer

Secondary antibodies

The light-harvesting protein Lhca4 is one of the four main and highly conserved types of chlorophyll a/b-binding proteins (Lhca1-4) of the light harvesting antenna (LHCI) of plant photosystem I. Lhca4 is imported as a precursor from the cytosol into the chloroplast. Upon insertion into the thylakoid membrane Lhca4 forms a heterodimer (LHCI-730) with Lhca1 that associates with the PSI core close to PsaG and PsaF.
A biochemical characterization of the plant LHCI antenna can be found in Klimmek et al. (2005) The structure of the higher plant light harvesting complex I: in vivo characterization and structural interdependence of the Lhca proteins. Biochemistry 44: 3065–3073.