Lhca2 | PSI type II chloropyll a/b-binding protein

AS01 006 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Monocots and dicots; A. thaliana, A. hypogaea, B. corticulans, C. reticulata, C. quitensis Kunt Bartl, C.reinhardti (one Lhca-type), C. zofingiensis,H. vulgare, N. tabacum, O. sativa, P. sativum, P. vulgaris, P. patens, P. banksiana, Prasinoderma sp., Pyramimonas sp., S. oleracea, Triticale, Z. mays

Lhca2 | PSI type II chloropyll a/b-binding protein

Product Information

Immunogen

BSA-conjugated synthetic peptide derived from the Lhca2 protein of Arabidopsis thaliana UniProt: Q9SYW8, Q8LCQ4, TAIR: At3g61470. This sequence is highly conserved in Lhca2 proteins of angiosperms (monocots and dicots) and gymnosperms as well as in At1g19150. This gene codes for the very low expressed Lhca6 protein which also has been denoted as Lhca2*1.

Host Rabbit

Clonality Polyclonal

Purity Total IgG. Protein G purified in PBS pH 7.4.

Format Lyophilized

Quantity 0.5 mg

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 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 : 2000-1 : 5000 (WB)

Expected | apparent MW

27.7 | 24 kDa for Arabidopsis thaliana

Reactivity

Confirmed reactivity Arabidopsis thaliana, Arachis hypogaea, Bryopsis corticulans, Colobanthus quitensis Kunt Bartl, Chlamydomonas reinhardti (one Lhca-type), Citrus reticulata, Chromochloris zofingiensis, Cytisus cantabricus (Wilk.) Rchb. F., Hieracium pilosella L, Hordeum vulgare, Lasallia hispanica, Nicotiana tabacum, Oryza sativa, Pisum sativum, Phaseolus vulgaris, Physcomitrium patens, Pinus banksiana (the higher of the two bands detected at 24 and 30 kDa is not considered to be specific to any Lhc protein), Posidonia oceanica, Prasinoderma sp., Pyramimonas sp. Spinacia oleracea, Syntrichia muralis (Hedw.) Raab, Triticum aestivum, Triticale, Zea mays

Predicted reactivity Dicots, Gymnosperms

Not reactive in No confirmed exceptions from predicted reactivity are currently known

Application examples

Application examples Application example

western blot using anti-Lhca2 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

Reactant: Nicotiana tabacum (Common tobacco)

Application: Western Blotting

Pudmed ID: 28180288

Journal: J Exp Bot

Figure Number: 5A

Published Date: 2017-02-01

First Author: Schöttler, M. A., Thiele, W., et al.

Impact Factor: 6.088

Open Publication

Immunoblot analysis of photosynthetic complex accumulation in wild-type tobacco and the two ?psaI lines grown under low, intermediate, and high-light conditions. Because the accumulation of most tested proteins was highest under high-light conditions, lanes one to three contain samples diluted to 25%, 50%, and a 100% sample of wild-type tobacco grown under high-light conditions, to allow for semi-quantitative determination of changes in protein abundance. Lanes four and five contain the two transplastomic lines grown at 1000 µE m?2 s?1. Lanes six to eight contain wild-type tobacco and the mutants grown at intermediate light intensities, and lanes nine to eleven contain samples grown at low light intensities. For PSII, the accumulation of the essential subunits PsbB (CP43) and PsbD (D2) and the LHCB1 antenna protein were determined, while for the cytochrome b6f complex, the accumulation of the essential redox-active subunits PetA (cytochrome f), PetB (cytochrome b6), and PETC (Rieske FeS protein) was tested. AtpB was probed as an essential subunit of the chloroplast ATP. For PSI, in addition to the three essential plastome-encoded subunits PsaA, PsaB, and PsaC, the accumulation of the nuclear-encoded subunits PSAD, PSAH, PSAK, PSAL, and PSAN and of the four LHCI proteins (LHCA1, LHCA2, LHCA3, LHCA4) was determined. Finally, we examined the accumulation of Ycf4, the chloroplast-encoded PSI-biogenesis factor encoded in the same operon as PsaI, and the nuclear-encoded assembly factor Y3IP1.

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.

Background

The light-harvesting protein Lhca2 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. Lhca2 is imported as a precursor from the cytosol into the chloroplast. Upon integration in the thylakoid membrane Lhca2 forms a heterodimer (LHCI-680) with Lhca3 that associates with the PSI core close to PsaF and PsaK.
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.

Product citations

Selected references Sarvari et al. (2022). Qualitative and quantitative evaluation of thylakoid complexes separated by Blue Native PAGE. Plant Methods. 2022 Mar 3;18(1):23. doi: 10.1186/s13007-022-00858-2. PMID: 35241118; PMCID: PMC8895881.
Fukura et al. (2021) Enrichment of chlorophyll catabolic enzymes in grana margins and their cooperation in catabolic reactions. J Plant Physiol. 2021 Nov;266:153535. doi: 10.1016/j.jplph.2021.153535. Epub 2021 Sep 25. PMID: 34607178.
Zhu et al. (2020). A NAC transcription factor and its interaction protein hinder abscisic acid biosynthesis by synergistically repressing NCED5 in Citrus reticulata. J Exp Bot. 2020 Jun 22;71(12):3613-3625.doi: 10.1093/jxb/eraa118.
Their et al. (2020). VIPP2 interacts with VIPP1 and HSP22E/F at chloroplast membranes and modulates a retrograde signal for HSP22E/F gene expression. Plant Cell Environ. 2020 Jan 29. doi: 10.1111/pce.13732.
Vojta and Fulgosi (2019). Topology of TROL protein in thylakoid membranes of Arabidopsis thaliana. Physiol Plant. 2019 Jan 20. doi: 10.1111/ppl.12927.
Roth et al. (2019). Regulation of Oxygenic Photosynthesis during Trophic Transitions in the Green Alga Chromochloris zofingiensis. Plant Cell. 2019 Feb 20. pii: tpc.00742.2018. doi: 10.1105/tpc.18.00742.
Mao et al. (2018). Comparison on Photosynthesis and Antioxidant Defense Systems in Wheat with Different Ploidy Levels and Octoploid Triticale. Int J Mol Sci. 2018 Oct 2;19(10). pii: E3006. doi: 10.3390/ijms19103006.
Kim et al. (2018). The rice zebra3 (z3) mutation disrupts citrate distribution and produces transverse dark-green/green variegation in mature leaves. Rice (N Y). 2018 Jan 5;11(1):1. doi: 10.1186/s12284-017-0196-8.
Myouga et al. (2018). Stable accumulation of photosystem II requires ONE-HELIX PROTEIN1 (OHP1) of the light harvesting-like family. Plant Physiol. 2018 Feb 1. pii: pp.01782.2017. doi: 10.1104/pp.17.01782.
Schottler et al. (2017). The plastid-encoded PsaI subunit stabilizes photosystem I during leaf senescence in tobacco. J Exp Bot. 2017 Feb 1;68(5):1137-1155. doi: 10.1093/jxb/erx009.
Chen et al. (2017). Comparison of Photosynthetic Characteristics and Antioxidant Systems in Different Wheat Strains. J Plant Growth Regul.
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.
Míguez et al. (2017). Diversity of winter photoinhibitory responses: A case study in co-occurring lichens, mosses, herbs and woody plants from subalpine environments. Physiol Plant. 2017 Feb 14. doi: 10.1111/ppl.12551.
Hu et al. (2017). The SUFBC2 D Complex is Required for the Biogenesis of All Major Classes of Plastid Fe-S Proteins. Plant J. 2017 Jan 19. doi: 10.1111/tpj.13483.
Kunugi et al. (2016). Evolution of Green Plants Accompanied Changes in Light-Harvesting Systems. Plant Cell Physiol. 2016 Jun;57(6):1231-43. doi: 10.1093/pcp/pcw071. Epub 2016 Apr 6.
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.

background:	The light-harvesting protein Lhca2 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. Lhca2 is imported as a precursor from the cytosol into the chloroplast. Upon integration in the thylakoid membrane Lhca2 forms a heterodimer (LHCI-680) with Lhca3 that associates with the PSI core close to PsaF and PsaK. 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.
More images:	Reactant: Nicotiana tabacum (Common tobacco) Application: Western Blotting Pudmed ID: 28180288 Journal: J Exp Bot Figure Number: 5A Published Date: 2017-02-01 First Author: Schöttler, M. A., Thiele, W., et al. Impact Factor: 6.088 Open Publication Immunoblot analysis of photosynthetic complex accumulation in wild-type tobacco and the two ?psaI lines grown under low, intermediate, and high-light conditions. Because the accumulation of most tested proteins was highest under high-light conditions, lanes one to three contain samples diluted to 25%, 50%, and a 100% sample of wild-type tobacco grown under high-light conditions, to allow for semi-quantitative determination of changes in protein abundance. Lanes four and five contain the two transplastomic lines grown at 1000 µE m?2 s?1. Lanes six to eight contain wild-type tobacco and the mutants grown at intermediate light intensities, and lanes nine to eleven contain samples grown at low light intensities. For PSII, the accumulation of the essential subunits PsbB (CP43) and PsbD (D2) and the LHCB1 antenna protein were determined, while for the cytochrome b6f complex, the accumulation of the essential redox-active subunits PetA (cytochrome f), PetB (cytochrome b6), and PETC (Rieske FeS protein) was tested. AtpB was probed as an essential subunit of the chloroplast ATP. For PSI, in addition to the three essential plastome-encoded subunits PsaA, PsaB, and PsaC, the accumulation of the nuclear-encoded subunits PSAD, PSAH, PSAK, PSAL, and PSAN and of the four LHCI proteins (LHCA1, LHCA2, LHCA3, LHCA4) was determined. Finally, we examined the accumulation of Ycf4, the chloroplast-encoded PSI-biogenesis factor encoded in the same operon as PsaI, and the nuclear-encoded assembly factor Y3IP1.
Picture (footer):	Application example 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
calculated \| apparent molecular mass [kDa]:	27.7 \| 24 kDa for Arabidopsis thaliana
Clonality:	Polyclonal
Format:	Lyophilized
Host:	Rabbit
immunogen:	BSA-conjugated synthetic peptide derived from the Lhca2 protein of Arabidopsis thaliana UniProt: Q9SYW8, Q8LCQ4, TAIR: At3g61470. This sequence is highly conserved in Lhca2 proteins of angiosperms (monocots and dicots) and gymnosperms as well as in At1g19150. This gene codes for the very low expressed Lhca6 protein which also has been denoted as Lhca2*1.
Purity:	Total IgG. Protein G purified in PBS pH 7.4.
Quantity:	0.5 mg
recommended dilution:	1 : 2000-1 : 5000 (WB)
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 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)
All references:	Sarvari et al. (2022). Qualitative and quantitative evaluation of thylakoid complexes separated by Blue Native PAGE. Plant Methods. 2022 Mar 3;18(1):23. doi: 10.1186/s13007-022-00858-2. PMID: 35241118; PMCID: PMC8895881. Fukura et al. (2021) Enrichment of chlorophyll catabolic enzymes in grana margins and their cooperation in catabolic reactions. J Plant Physiol. 2021 Nov;266:153535. doi: 10.1016/j.jplph.2021.153535. Epub 2021 Sep 25. PMID: 34607178. Zhu et al. (2020). A NAC transcription factor and its interaction protein hinder abscisic acid biosynthesis by synergistically repressing NCED5 in Citrus reticulata. J Exp Bot. 2020 Jun 22;71(12):3613-3625.doi: 10.1093/jxb/eraa118. Their et al. (2020). VIPP2 interacts with VIPP1 and HSP22E/F at chloroplast membranes and modulates a retrograde signal for HSP22E/F gene expression. Plant Cell Environ. 2020 Jan 29. doi: 10.1111/pce.13732. Vojta and Fulgosi (2019). Topology of TROL protein in thylakoid membranes of Arabidopsis thaliana. Physiol Plant. 2019 Jan 20. doi: 10.1111/ppl.12927. Roth et al. (2019). Regulation of Oxygenic Photosynthesis during Trophic Transitions in the Green Alga Chromochloris zofingiensis. Plant Cell. 2019 Feb 20. pii: tpc.00742.2018. doi: 10.1105/tpc.18.00742. Mao et al. (2018). Comparison on Photosynthesis and Antioxidant Defense Systems in Wheat with Different Ploidy Levels and Octoploid Triticale. Int J Mol Sci. 2018 Oct 2;19(10). pii: E3006. doi: 10.3390/ijms19103006. Kim et al. (2018). The rice zebra3 (z3) mutation disrupts citrate distribution and produces transverse dark-green/green variegation in mature leaves. Rice (N Y). 2018 Jan 5;11(1):1. doi: 10.1186/s12284-017-0196-8. Myouga et al. (2018). Stable accumulation of photosystem II requires ONE-HELIX PROTEIN1 (OHP1) of the light harvesting-like family. Plant Physiol. 2018 Feb 1. pii: pp.01782.2017. doi: 10.1104/pp.17.01782. Schottler et al. (2017). The plastid-encoded PsaI subunit stabilizes photosystem I during leaf senescence in tobacco. J Exp Bot. 2017 Feb 1;68(5):1137-1155. doi: 10.1093/jxb/erx009. Chen et al. (2017). Comparison of Photosynthetic Characteristics and Antioxidant Systems in Different Wheat Strains. J Plant Growth Regul. 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. Míguez et al. (2017). Diversity of winter photoinhibitory responses: A case study in co-occurring lichens, mosses, herbs and woody plants from subalpine environments. Physiol Plant. 2017 Feb 14. doi: 10.1111/ppl.12551. Hu et al. (2017). The SUFBC2 D Complex is Required for the Biogenesis of All Major Classes of Plastid Fe-S Proteins. Plant J. 2017 Jan 19. doi: 10.1111/tpj.13483. Kunugi et al. (2016). Evolution of Green Plants Accompanied Changes in Light-Harvesting Systems. Plant Cell Physiol. 2016 Jun;57(6):1231-43. doi: 10.1093/pcp/pcw071. Epub 2016 Apr 6. 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.
Confirmed reactivity:	Arabidopsis thaliana, Arachis hypogaea, Bryopsis corticulans, Colobanthus quitensis Kunt Bartl, Chlamydomonas reinhardti (one Lhca-type), Citrus reticulata, Chromochloris zofingiensis, Cytisus cantabricus (Wilk.) Rchb. F., Hieracium pilosella L, Hordeum vulgare, Lasallia hispanica, Nicotiana tabacum, Oryza sativa, Pisum sativum, Phaseolus vulgaris, Physcomitrium patens, Pinus banksiana (the higher of the two bands detected at 24 and 30 kDa is not considered to be specific to any Lhc protein), Posidonia oceanica, Prasinoderma sp., Pyramimonas sp. Spinacia oleracea, Syntrichia muralis (Hedw.) Raab, Triticum aestivum, Triticale, Zea mays
predicted reactivity:	Dicots, Gymnosperms
not reactive in:	No confirmed exceptions from predicted reactivity are currently known
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.

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