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Lhcb2 | LHCII type II chlorophyll a/b-binding protein

AS01 003  |  Clonality: Polyclonal  |  Host: Rabbit  |  Reactivity: Photosynthetic eukaryotes including A. thaliana, A. hypogaea, B. sylvaticum, , C. arietinum, C. quitensis Kunt Bartl, C. sativa, H. vulgare, C. reinhardtii, L. esculentum (Solanum lycopersicon), M. crystallinum, N. tabacum, O. sativa, P. patens, P. sativum, P. vulgaris, S. alba, S. oleracea, T. aestivum, Triticale, Z. mays

Lhcb2 | LHCII type II chlorophyll a/b-binding protein in the group Antibodies Plant/Algal  / Photosynthesis  / LHC at Agrisera AB (Antibodies for research) (AS01 003)
Lhcb2 | LHCII type II chlorophyll a/b-binding protein



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Product Information

Immunogen

BSA-conjugated synthetic peptide derived from a highly conserved sequence of Lhcb2 proteins from angiosperms (monocots and dicots) and gymnosperms, including Arabidopsis thaliana Lhcb2.1 UniProt: Q9SHR7, TAIR: AT2G05100, Lhcb2.2 UniProt: Q9S7J7, TAIR:AT2G05070, Lhcb2.3 UniProt:Q9XF87, TAIR:AT3G27690
 

Host Rabbit
Clonality Polyclonal
Purity Immunogen affinity purified serum in PBS pH 7.4.
Format Lyophilized
Quantity 50 ĩg
Reconstitution For reconstitution add 50 ĩl 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 Immunoprecipitation (IP), ImmunoGold (IG), Western blot (WB)
Recommended dilution 5 µl of antibody solution (IP), 1: 100 (IG), 1: 500 - 1 : 5000 (WB)
Expected | apparent MW

28.6 | 25 kDa for Arabidopsis thaliana

Reactivity

Confirmed reactivity Acer pseudoplatanus, Arabidopsis thaliana, Arachis hypogaea, Brachypodium sylvaticum, Cicer arietum, Chlorella vulgaris, Colobanthus quitensis Kunt Bartl, Chlamydomonas reinhardtii, Cucumis sativus, Cytisus cantabricus (Wilk.) Rchb. F., Hieracium pilosella L., Hieracium pilosella L., Hordeum vulgare, Lasallia hispanica, Lycopersicon esculentum (Solanum lycopersicon), Miscanthus x giganteus, Mesembryanthemum crystallinum, Nicotiana benthamiana, Nicotiana tabacum, Oryza sativa, Pisum sativum, Phaseolus coccineus L., Phaseolus vulgaris,Physcomitrium patens, Sinapsis alba, Spinacia oleracea, Syntrichia muralis (Hedw.) Raab, Triticum aestivum, Triticale, Zea mays
Predicted reactivity Algae, Dicots, Gymnosperms, Mosses
Not reactive in No confirmed exceptions from predicted reactivity are currently known

Application examples

Application examples Application example

Western blot using anti-Lhcb2 antibodies

Species and variants: Pea – Pisum sativum L. Bean – Phaseolus coccineus L. 3h – 3 hours of cold exposure 9h – 9 hours of cold exposure 12h – 12 hours of cold exposure 2d – 2 days of cold exposure

Samples of isolated thylakoids containing 3 µg of chlorophyll were denatured with Laemmli buffer (1 vol : 1 vol) at 75 °C for 5 min. Denatured samples containing 1 µg of chlorophyll were loaded in the gel wells, separated on 12% SDS-PAGE gels and blotted for 45 min at 100 V to PVDF membrane using wet transfer. Blot was blocked with 5% milk in TBS-T for 60 min at room temperature (RT) with agitation. The blot was incubated with the primary antibody at a dilution of 1:500 in 1% Amersham™ ECL Prime Blocking Agent in TBS-T overnight at 4ºC with agitation. The antibody solution was decanted and the blot was washed 3 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: 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, 1 time for 5 min in TBS, 1 time for 5 min in 0.1 M Tris (pH 8.5), and developed for 4 min in substrates (0.188 mM coumaric acid, 1.25 mM luminol, 0.01% H2O2). Exposure time was 5 seconds in ChemiDoc scanner (BioRad).

Msc Małgorzata Krysiak, Faculty of Biology, University of Warsaw, Poland

Reactant: Plant

Application: Western Blotting

Pudmed ID: 27590049

Journal: BMC Plant Biol

Figure Number: 9A

Published Date: 2016-09-02

First Author: Mazur, R., Sadowska, M., et al.

Impact Factor: 4.142

Open Publication

Changes of PSII and PSI antenna and core protein levels. Proteins from control and Tl-treated white mustard leaves were separated by SDS-PAGE followed by immunodetection with antibodies against Lhcb1, Lhcb2, Lhca1 (antenna proteins) and D1, D2, CP43, PsbO, PsaA (core proteins). Samples were loaded on the equal amount of chlorophyll (0.25 ?g). Description of samples abbreviation as given in the legend to Fig. 3


Reactant: Arabidopsis thaliana (Thale cress)

Application: Western Blotting

Pudmed ID: 31240258

Journal: Commun Biol

Figure Number: 2b

Published Date: 2019-06-27

First Author: Pralon, T., Shanmugabalaji, V., et al.

Impact Factor: None

Open Publication

Thylakoid protein phosphorylation and state transitions are disturbed after high light treatment in pgr6 background. a Total protein extracts of 4-week-old wild type (WT), pgr6-1, pgr6?2, sps2 and stn7/stn8 analysed by immunoblotting with anti-phosphothreonine antibody; the principal thylakoid phospho proteins are indicated on the right according to their size. Core photosystem II proteins D1 (PsbA) and D2 (PsbD) are indicated as a single band due to their poor resolution. Actin was used as a loading control. b Lhcb1 and Lhcb2 phosphorylation levels were visualised after separation on Phostag™-pendant acrylamide gels. The upper band corresponds to the phosphorylated form (p-), stn7/stn8 double mutant is a non-phosphorylated control. c Average transient of the variable room temperature chlorophyll fluorescence measured during the transition from red (660nm) supplemented with far-red light (720nm) state 1 to pure red light state 2 (n?=?4 independent pots containing 2–3 plants). The fluorescence curves from pgr6 and sps2 are shifted on the x-axis to allow visualising the FMST1 and FMST2 values. The x-axis time scale refers to the wild-type curve. d Calculated quenching related to state transition (qT?=?(FMST1?–?FMST2)/FM), expressed as the percentage of FM that is dissipated by the state 1 to state 2 transition, of wild type (WT), pgr6?1 and sps2 under moderate light (120?mol?m?2?s?1) (ML) and after 3?h of high light (500?mol?m?2?s?1) (HL). Whiskers and box plot shows the minimum, first quartile, median, average, third quartile and maximum of each dataset (n?=?4 biologically independent samples); p-values are calculated via a two-tailed Student’s t test. e STN7 phosphorylation level visualised after separation on Phostag™-pendant acrylamide gels. The upper band corresponds to the phosphorylated form (p-), a protein sample from stn7/stn8 double mutant was loaded as a control for the antibody specificity. Uncropped images of the membranes displayed in a, b and e are available as Supplementary Fig. 11. Data points for items c, d are available as Supplementary data 2


Reactant: Arabidopsis thaliana (Thale cress)

Application: Western Blotting

Pudmed ID: 32003746

Journal: Elife

Figure Number: 8C

Published Date: 2020-01-31

First Author: Cazzonelli, C. I., Hou, X., et al.

Impact Factor: 7.448

Open Publication

Chemical inhibition of CCD activity revealed how a ccr2 generated apocarotenoid signal transcriptionally represses HY5 and LHCB2 expression during photomorphogenesis.(A) Transcript levels of PIF3 and HY5 in WT, ccr2, ccr2 det1-154 and det1-154 de-etiolated seedlings growing on MS media + /- D15. (B) Representative western blot images showing PIF3 and HY5 protein levels in WT, ccr2, ccr2 det1-154 and det1-154 de-etiolated seedlings growing on MS media + /- D15. The membrane was re-probed using anti-Actin antibody as an internal loading control. (C) Protein and transcript levels of LHCB2 expression in WT and ccr2 de-etiolated seedlings growing on MS media + /- D15. (D) Model showing how ACS regulates HY5 and LHCB2 expression in ccr2. Images of seedlings represent are cotyledons are coloured green or yellow to reflect the delay in chlorophyll biosynthesis induced by ACS as evidenced in Figure 6c. De-etiolation of seedlings was performed by transferring 4-d-old etiolated seedlings to continuous light for 3 d to induce photomorphogenesis. Statistical analysis denoted as a star was performed by pair-wise t-test (p<0.05). Error bars represent standard error of means. Ctrl; Control; Ctrl, D15; chemical inhibitor of CCD activity.


Reactant: Arabidopsis thaliana (Thale cress)

Application: Western Blotting

Pudmed ID: 32269582

Journal: Front Plant Sci

Figure Number: 7A,B

Published Date: 2020-04-10

First Author: Pralon, T., Collombat, J., et al.

Impact Factor: 5.435

Open Publication

Double mutant maintains thylakoid protein phosphorylation and state transitions after high light. (A) Total protein extracts of wild type (WT), abc1k1.1, -2, abc1k3.1, -2, and abc1k1/abc1k3.1, -2 light-exposed leaves were separated by SDS PAGE, transferred on nitrocellulose membrane and decorated with anti-phosphothreonine antibody. The main thylakoid phospho-proteins are indicated on the right according to their size. Core photosystem II proteins D1 (PsbA) and D2 (PsbD) are indicated together due their poor resolution. (B) The accumulation of the principal photosynthetic complexes was assessed using antibodies against specific subunits of each complex: anti-Lhcb2 for the major LHCII, anti-D1 (PsbA) for PSII, anti-PetC for cytochrome b6f, anti-PsaD and anti-PsaC for PSI, and anti-AtpC for ATP synthase. Actin signal is shown as a loading control. (C) Fluorescence quenching related to the state transitions (qT) of wild type (WT), abc1k1.1, -2, abc1k3.1, -2, and abc1k1/abc1k3.1, -2 under moderate light (120 ?mol of photons m–2 s–1) (ML) and after 3 h of high light (500 ?mol of photons m–2 s–1) (HL). qT was calculated from the maximal chlorophyll fluorescence measured after 10 min exposure to red light (660 nm) supplemented with far-red illumination (720 nm) “State 1” (FMST1) or to pure red light “State 2” (FMST2). Quenching related to state transition was calculated as qT = (FMST1 – FMST2)/FM. Each value represents the average of a pot containing 2–3 plants. Superscript letters are used to indicate statistically different groups (p < 0.05) by paired Student’s t-test.


Reactant: Arabidopsis thaliana (Thale cress)

Application: Western Blotting

Pudmed ID: 33322466

Journal: Biomolecules

Figure Number: 7A

Published Date: 2020-12-11

First Author: Andreeva, A. A., Vankova, R., et al.

Impact Factor: None

Open Publication

Immunoblot analysis of the photosynthetic proteins on the basis of equal total Ponceau S dye stained blot with proteins from leaves of wild type plants and pap1 mutant grown on MS medium in Petri dishes for four weeks under a 16 h light/8 h dark photoperiod at 23 °C with 100 ?E m?2 s?1. Proteins were visualized by immunoblotting using antibodies specific for RbcL, PsaB, PsbD, AtpB, RpoB, AccD and Lhcb2.4 proteins.


Reactant: Arabidopsis thaliana (Thale cress)

Application: Western Blotting

Pudmed ID: 33629953

Journal: Elife

Figure Number: 6A

Published Date: 2021-02-25

First Author: Pipitone, R., Eicke, S., et al.

Impact Factor: 7.448

Open Publication

Accumulation dynamics of photosynthesis-related proteins during de-etiolation.Three-day-old etiolated seedlings of Arabidopsis thaliana were illuminated for 0 hr (T0), 4 hr (T4), 8 hr (T8), 12 hr (T12), 24 hr (T24), 48 hr (T48), 72 hr (T72), and 96 hr (T96) under white light (40 ĩmol/m2/s). (A) Proteins were separated by SDS-PAGE and transferred onto nitrocellulose membrane and immunodetected with antibodies against PsbA, PsbD, PsbO, PetC, PsaD, PsaC, Lhcb2, AtpC, ELIP, POR, phyA, HY5, and ACTIN proteins. (B–C) Quantification of PsbA, PetC, and PsaC during de-etiolation. Heatmap (B) was generated after normalization of the amount of each protein relative to the last time point (T96). Graph (C) corresponds to the absolute quantification of proteins at T96. Error bars indicate ą SD (n = 3). Quantification of photosystem-related proteins during de-etiolation is detailed in Figure 6—figure supplement 1.Figure 6—source data 1.Quantitative data for immunoblot analysis.Quantitative data for immunoblot analysis.Quantification of photosynthesis-related proteins.(A) Immunodetection of PsbA, PetC, and PsaC during de-etiolation. Dilutions were used for the later time points to avoid saturation of the signal. (B) Different bands were detected by Amersham Imager program and quantified by Image QuantTL (Amersham). (C) Calibration curves were created using recombinant proteins (Agrisera). Calibration curve composition: PsbA 10 ng (A; lane a), 5 ng (b), 2.5 ng (c), and 1.25 ng (d); PetC 10 ng (e), 5 ng (f), 2.5 ng (g), and 1.25 ng (h); PsaC 3 ng (i), 1.5 ng (l), 0.75 ng (m), and 0.325 ng (n). Data indicate mean ą SD (n = 3–4). Raw data and calculations are shown in Figure 6—source data 1.

Additional information

Immunoprecipitation has been done using Immunoprecipitation kit from Roche, Cat.No. 11 719 386 001.

Protein is processed into mature form (Jansson 1999).

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Plant protein extraction buffer

Background

Background

The major light-harvesting antenna complex II (LHCII) in photosynthetic eukaryotes is located in the thylakoid membrane of the chloroplast. It is a heterotrimeric complex formed by up to 3 different individual subtypes of chlorophyll a/b-binding proteins: Lhcb1, Lhcb2, and Lhcb3. Lhcb2 is often coded by several nuclear genes and is found together with Lhcb1 within the mobile LHCII trimers involved in state1-state2 transition.
A molecular characterisation of the LHCII proteins can be found in Caffarri et al. (2004) A Look within LHCII:  Differential Analysis of the Lhcb1−3 Complexes Building the Major Trimeric Antenna Complex of Higher-Plant Photosynthesis. Biochemistry 43 (29): 9467–9476.

Product citations

Selected references Lande et al. (2022) Dehydration-responsive chickpea chloroplast protein, CaPDZ1, confers dehydration tolerance by improving photosynthesis. Physiologia Plantarum, 174( 1), e13613. Available from: https://doi.org/10.1111/ppl.13613
Pipitone et al. (2021). A multifaceted analysis reveals two distinct phases of chloroplast biogenesis during de-etiolation in Arabidopsis. Elife. 2021 Feb 25;10:e62709. doi: 10.7554/eLife.62709. PMID: 33629953; PMCID: PMC7906606.
Kamea et al. (2021). Substitution of deoxycholate with the amphiphilic polymer amphipol A8-35 improves the stability of large protein complexes during native electrophoresis. Plant Cell Physiol. 2021 Jan 5:pcaa165. doi: 10.1093/pcp/pcaa165. Epub ahead of print. PMID: 33399873.
Wu et al. (2021). Formation of light-harvesting complex (LHC) II aggregates from LHCII-PSI-LHCI complexes in rice plants under high light. J Exp Bot. 2021 May 3:erab188. doi: 10.1093/jxb/erab188. Epub ahead of print. PMID: 33939808.
Mazur et al. (2021) The SnRK2.10 kinase mitigates the adverse effects of salinity by protecting photosynthetic machinery. Plant Physiol. 2021 Dec 4;187(4):2785-2802. doi: 10.1093/plphys/kiab438. PMID: 34632500; PMCID: PMC8644180.
Pavlovic & Kocab. (2021) Alternative oxidase (AOX) in the carnivorous pitcher plants of the genus Nepenthes: what is it good for? Ann Bot. 2021 Dec 18:mcab151. doi: 10.1093/aob/mcab151. Epub ahead of print. PMID: 34922341.
Toubiana et al. (2020). Correlation-based Network Analysis Combined With Machine Learning Techniques Highlight the Role of the GABA Shunt in Brachypodium Sylvaticum Freezing Tolerance. Sci Rep , 10 (1), 4489
Grieco et al. (2020). Adjustment of photosynthetic activity to drought and fluctuating light in wheat. Plant Cell Environ. 2020 Mar 16. doi: 10.1111/pce.13756.
Hertle et al. (2020) A Sec14 Domain Protein Is Required for Photoautotrophic Growth and Chloroplast Vesicle Formation in Arabidopsis thaliana. Proc Natl Acad Sci USA 2020 Apr 3 (Immunogold)
Pralon et al. (2019). Plastoquinone homoeostasis by Arabidopsis proton gradient regulation 6 is essential for photosynthetic efficiency. Commun Biol. 2019 Jun 20;2:220. doi: 10.1038/s42003-019-0477-4.
Lv et al. (2019). Uncoupled Expression of Nuclear and Plastid Photosynthesis-Associated Genes Contributes to Cell Death in a Lesion Mimic Mutant. Plant Cell. 2019 Jan;31(1):210-230. doi: 10.1105/tpc.18.00813.
Rogowski et al. (2019). Photosynthesis and organization of maize mesophyll and bundle sheath thylakoids of plants grown in various light intensities. Environmental and Experimental Botany Volume 162, June 2019, Pages 72-86.
Koh et al. (2019). Heterologous synthesis of chlorophyll b in Nannochloropsis salina enhances growth and lipid production by increasing photosynthetic efficiency. Biotechnol Biofuels. 2019 May 14;12:122. doi: 10.1186/s13068-019-1462-3. eCollection 2019.
Bethmann et al. (2019). The zeaxanthin epoxidase is degraded along with the D1 protein during photoinhibition of photosystem II. Plant Direct. 2019 Dec 1;3(11):e00185. doi: 10.1002/pld3.185.
Gayen et al. (2018). Dehydration-induced proteomic landscape of mitochondria in chickpea reveals large-scale coordination of key biological processes. J Proteomics. 2018 Sep 19. pii: S1874-3919(18)30349-X. doi: 10.1016/j.jprot.2018.09.008
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.
Tadini et al. (2018). Trans-splicing of plastid rps12 transcripts, mediated by AtPPR4, is essential for embryo patterning in Arabidopsis thaliana. Planta. 2018 Jul;248(1):257-265. doi: 10.1007/s00425-018-2896-8.
Li et al. (2018). Modulating plant growth-metabolism coordination for sustainable agriculture. Nature. 2018 Aug 15. doi: 10.1038/s41586-018-0415-5.
Shanmugabalaji et al. (2018). Chloroplast Biogenesis Controlled by DELLA-TOC159 Interaction in Early Plant Development. Curr Biol. 2018 Aug 20;28(16):2616-2623.e5. doi: 10.1016/j.cub.2018.06.006.
Du et al. (2018). Galactoglycerolipid Lipase PGD1 Is Involved in Thylakoid Membrane Remodeling in Response to Adverse Environmental Conditions in Chlamydomonas. Plant Cell. 2018 Feb;30(2):447-465. doi: 10.1105/tpc.17.00446.
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.
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.
Rantala et al. (2017). Proteomic characterization of hierarchical megacomplex formation in Arabidopsis thylakoid membrane. Plant J. 2017 Dec;92(5):951-962. doi: 10.1111/tpj.13732.
Shin et al. (2017), Complementation of a mutation in CpSRP43 causing partial truncation of light-harvesting chlorophyll antenna in Chlorella vulgaris. Sci Rep. 2017 Dec 20;7(1):17929. doi: 10.1038/s41598-017-18221-0.
Cantrell and Peers (2017). A mutant of Chlamydomonas without LHCSR maintains high rates of photosynthesis, but has reduced cell division rates in sinusoidal light conditions. PLoS One. 2017 Jun 23;12(6):e0179395. doi: 10.1371/journal.pone.0179395.
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)
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.
Yang-Er Chen et al. (2017). Responses of photosystem II and antioxidative systems to high light and high temperature co-stress in wheat. J. of Exp. Botany, Volume 135, March 2017, Pages 45–55.

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