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Agrisera Super Deal

Primary/Secondary/ECL



Add only 20 € to your primary antibody purchase
and you will also receive:

- Secondary antibody
Goat anti-rabbit, HRP conjugated
(to be used >1: 25 000)


- Two chemiluminescent detection reagents
Agrisera ECL Bright/SuperBright,  for 10 midi blots (picogram and femtogram detection range)

AS18 PrimarySecondaryECL

PsbB | CP47 protein of PSII

AS04 038 | Clonality: Polyclonal | Host: Rabbit | Reactivity: [global antibody] for higher plants, Physcomitrella patens, algae, cyanobacteria, diatoms

PsbB | CP47 protein of PSII in the group Plant/Algal Antibodies / Global Antibodies at Agrisera AB (Antibodies for research) (AS04 038)

PRODUCT INFORMATION IN PDF

Qty: 
360 €

Info: Read reviews
product information
Background

PsbB (CP47) is a chlorophyll-binding protein located in the membrane, where it serves as the core antenna of Photosystem II.

Immunogen

KLH-conjugated synthetic peptide derived from available plant, algal and cyanobacterial PsbB sequences including Arabidopsis thaliana AtCg00680, Hordeum vulgare P10900, Oryza sativa P0C364, Synechocystis PCC 6803 P05429

Host Rabbit
Clonality Polyclonal
Clone
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)
Related products

AS04 038S | PsbB protein standard for quantitation and positive control is discontinued. We recommend to use PsbD | D2 together with PsbD antibody

AS04 038PRE | PsbB | CP47 protein of PSII, pre-immune serum
antibodies to other PSII proteins

recommended secondary antibody

Plant and algal protein extraction buffer

Secondary antibodies

Additional information This antibody can be used as a loading control for studies of PSIi or photosynthetic acclimation in diatoms Blommaert et al. 2017.  Limnol. Oceanogr. DOI: 10.1002/lno.10511.

This product can be sold containing ProClin if requested.
application information
Recommended dilution 1 : 2000 (WB)
Expected | apparent MW

56 kDa

Confirmed reactivity Anabaena 7120, Arabidopsis thaliana, Chlamydomonas reinhardtii, Echinochloa crus-galli, Hordeum vulgare, Malus prunifolia, Opephora guenter-grassii (diatom), Oryza sativa, Panicum miliaceum, Phaseolus vulgaris, Physcomitrella patens, Pisum sativum, Synechococcus PCC7942, 6803, , Seminavis robusta (diatom), Zea mays
Predicted reactivity Abies concolor, Brachypodum distachyon, Brassica napus, Cannabis sativa, Cyanobacteria, Cucumis sativus, Ephedra sp., Glycine max, Lotus japonicus, Nanochloropsis sp. , Nicotiana tabacum, Panax ginseng, Populus trichocarpa, Solanum tuberosum, Sorghum bicolor, Spinacia oleracea, Triticum aestivum, Vitis vinifera
Not reactive in No confirmed exceptions from predicted reactivity are currently known.
Additional information

This product can be sold containing ProClin if requested

in bis-tris gel systems PsbB protein migrates between 40-45 kDa

Selected references Bressan et al. (2018). Light harvesting complex I is essential for Photosystem II photoprotection under variable light conditions in Arabidopsis thaliana. Environmental and Experimental Botany Available online 10 March 2018.
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. Xing et al. (2017). Deletion of CGLD1 Impairs PSII and Increases Singlet Oxygen Tolerance of Green Alga Chlamydomonas reinhardtii. Front. Plant Sci., 15 December 2017.
Blommaert et al. (2017). Contrasting NPQ dynamics and xanthophyll cycling in a motile and a non-motile intertidal benthic diatom. Limnol. Oceanogr. doi: 10.1002/lno.10511
Gandini et al. (2017). The transporter SynPAM71 is located in the plasma membrane and thylakoids, and mediates manganese tolerance in Synechocystis PCC6803. New Phytol. 2017 Mar 20. doi: 10.1111/nph.14526.
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.
Fan et al. (2016). Proteome Analyses Using iTRAQ Labeling Reveal Critical Mechanisms in Alternate Bearing Malus prunifolia. J Proteome Res. 2016 Oct 7;15(10):3602-3616.

Application example

2 µg of total protein from (1) Arabidopsis thaliana leaf extracted with PEB (AS08 300), (2) Horderum vulgare leaf extracted with PEB (AS08 300), (3) Chlamydomonas reinhardtii total cell extracted with PEB (AS08 300), (4) Synechococcus sp. 7942 total cell extracted with PEB (AS08 300), (5) Anabaena sp. total cell extracted with PEB (AS08 300) were separated on 4-12% NuPage (Invitrogen) LDS-PAGE and blotted 1h to PVDF. Blots were blocked immediately following transfer in 2% ECL Advance blocking reagent (GE Healthcare) in 20 mM Tris, 137 mM sodium chloride pH 7.6 with 0.1% (v/v) Tween-20 (TBS-T) for 1h at room temperature with agitation. Blots were incubated in the primary antibody at a dilution of 1: 50 000 for 1h at room temperature with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed once for 15 min and 3 times for 5 min in TBS-T at room temperature with agitation. Blots were incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, recommended secondary antibody AS09 602) diluted to 1:50 000 in 2% ECL Advance blocking solution for 1h at room temperature with agitation. The blots were washed as above and developed for 5 min with ECL Advance detection reagent according the manufacturers instructions. Images of the blots were obtained using a CCD imager (FluorSMax, Bio-Rad) and Quantity One software (Bio-Rad).

 

Western blot of anti-PsbB antibody

application example

0.2 µg of chlorophyll from (4,5) Arabidopsis thaliana leaf extracted with PEB (AS08 300), (1) 500 fmol of PsbB protein standard, (2) 200 fmol of PsbB protein standard, (3) 75 fmol of PsbB protein standard were separated on 4-12% NuPage (Invitrogen) LDS-PAGE and blotted 1h to PVDF. Blots were blocked immediately following transfer in 2% ECL Advance blocking reagent (GE Healthcare) in 20 mM Tris, 137 mM sodium chloride pH 7.6 with 0.1% (v/v) Tween-20 (TBS-T) for 1h at room temperature with agitation. Blots were incubated in the primary antibody at a dilution of 1: 50 000 for 1h at room temperature with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed once for 15 min and 3 times for 5 min in TBS-T at room temperature with agitation. Blots were incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1:50 000 in 2% ECL Advance blocking solution for 1h at room temperature with agitation. The blots were washed as above and developed for 5 min with ECL Advance detection reagent according the manufacturers instructions. Images of the blots were obtained using a CCD imager (FluorSMax, Bio-Rad) and Quantity One software (Bio-Rad).

 

western blot detection using anti-PsbB antibody


Western blot using anti-CP47 antibodies

2.0 µg of chlorophyll from Pisum sativum chloroplasts and from Zea mays, Echinochloa crus-galli, Panicum miliaceum mesophyll and bundle sheath chloroplasts 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°C 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 in TBS for 2h at room temperature (RT) with agitation. Blot was incubated in the primary antibody AS04 038 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) 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 122 seconds.

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

||| For other applications, usage on species other than stated above or any other questions, please use the LiveChat option or contact us at support@agrisera.com