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Lhca1 | PSI type I chlorophyll a/b-binding protein

325 €

AS01 005  |  clonality: polyclonal  |  host: rabbit  |  reactivity: monocots and dicots including A. thaliana, A. hypogaea, C. quitensis Kunt Bartl, E. crus-galli,  G. hybrid, H. vulgare, L. esculentum (Solanum lycopersicum), N. tabacum, O. sativa, P. miliaceum, P. patens, P. sativum, P. strobus, P.vulgaris, S. oleracea, Z. mays

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AS01 005

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product information
Background

The light-harvesting protein Lhca1 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. Lhca1 is imported as a precursor from the cytosol into the chloroplast. Upon insertion into the thylakoid membrane Lhca1 forms a heterodimer (LHCI-730) with Lhca4 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

Immunogen

BSA-conjugated synthetic peptide derived from the Lhca1 protein of Arabidopsis thaliana UniProt: Q01667, TAIR:   At3g54890. This sequence is highly conserved in Lhca1 proteins of angiosperms (monocots and dicots) and gymnosperms.

Host Rabbit
Clonality Polyclonal
Clone
Purity Total IgG
Format Lyophilized in PBS pH 7.4.
Quantity

0.5mg

Reconstitution For reconstitution add 100 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)
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Available antibodies against pigment-binding proteins- LHC

recommended secondary antibody

Plant protein extraction buffer

Secondary antibodies

Additional information Antibody format is a total IgG fraction, which means that it is a pool of polyclonal antibodies obtained by purification of serum on Protein G, not on a specific antigen column.
application information
Recommended dilution

1 : 2000 - 1 : 5000 (WB)

Expected | apparent MW

25.99 | 22 kDa for Arabidopsis thaliana

Confirmed reactivity Arabidopsis thaliana, Arachis hypogaea, Bryopsis corticulans, Colobanthus quitensis Kunt Bartl, Echinochloa crus-galli, Guzmania hybrid, Hordeum vulgare, Lycopersicon esculentum (Solanum lycopersicum), Nicotiana tabacum, Oryza sativa, Panicum miliaceum, Physcomitrella patens, Pinus strobus, Pisum sativum, Phaseolus vulgaris, Spinacia oleracea, Zea mays,
Predicted reactivity

angiosperms (monocots and dicots), gymnosperms

Not reactive in

no confirmed exceptions from predicted reactivity known in the moment

Additional information Protein is processed into mature form (Jansson 1999).
Selected references Yang et al. (2017). Tetratricopeptide repeat protein Pyg7 is essential for photosystem I assembly by interacting with PsaC in Arabidopsis. Plant J. 2017 Jun 21. doi: 10.1111/tpj.13618.
Tyuereva et al. (2017). The absence of chlorophyll b affects lateral mobility of photosynthetic complexes and lipids in grana membranes of Arabidopsis and barley chlorina mutants. Photosynth Res. 2017 Apr 5. doi: 10.1007/s11120-017-0376-9. (Hordeum vulgare, western blot)
Mazur et al. (2016). Overlapping toxic effect of long term thallium exposure on white mustard (Sinapis alba L.) photosynthetic activity. BMC Plant Biol. 2016 Sep 2;16(1):191. doi: 10.1186/s12870-016-0883-4.
Heinnickel et al. (2016). Tetratricopeptide repeat protein protects photosystem I from oxidative disruption during assembly. Proc Natl Acad Sci U S A. 2016 Mar 8;113(10):2774-9. doi: 10.1073/pnas.1524040113
Qin et al. (2014). Isolation and characterization of a PSI-LHCI super-complex and its sub-complexes from a siphonaceous marine green alga, Bryopsis Corticulans. Photosynth Res. 2014 Sep 12.
Saito et al. (2014). Fe deficiency induces phosphorylation and translocation of Lhcb1 in barley thylakoid membranes. FEBS Lett. 2014 May 8. pii: S0014-5793(14)00317-2. doi: 10.1016/j.febslet.2014.04.031.

Application example 

western blot using anti-Lhca1 antibodies

1.0 µg of chlorophyll from mesophyll (M) and bundle sheath (BS) thylakoids of various C4 plants (Echinochloa crus-galli, Panicum miliaceum, Zea mays) extracted with 0.4 M sorbitol, 50 mM Hepes NaOH, pH 7.8, 10 mM NaCl, 5 mM MgCl2 and 2 mM EDTA. 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 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 2000 overnight at 4°C with agitation in 1% milk in TBS-T. 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 horse radish peroxidase conjugated, from Agrisera, AS09 602, Lot 1702) diluted to 1:25 000 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 15 seconds.

Courtesy of Dr. Wioleta Wasilewska, Warsaw University, Poland

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