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STN8 | Serine/threonine-protein kinase STN8 (chloroplastic)

AS10 1601 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana, Triticum aestivum

STN8 | Serine/threonine-protein kinase STN8 (chloroplastic) in the group Antibodies for Plant/Algal / Photosynthesis  / Kinases at Agrisera AB (Antibodies for research) (AS10 1601)

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product information
Background Serine/threonine protein kinase (EC=2.7.11.1), known as STN8 in Arabidopsis, and its Stl1 homologue in Chlamydomonas, are STN7/Stt7-like proteins that are not required for state transitions. The Arabidopsis mutants deficient in stn8 gene demonstrated highly reduced levels of the phosphorylated proteins of the photosystem II (Bonardi et al., 2005 Varionen et al., 2005). Alternative name: Protein STATE TRANSITION 8.
Immunogen KLH-conugated synthetic peptide (amino acids 425 - 438) specific for Arabidopsis thaliana STN8 serine/threonine proteinkinase, UniProt: Q9LZV4 TAIR:At5g01920
Host Rabbit
Clonality Polyclonal
Clone
Purity Affinity purified serum in PBS, pH 7.4
Format Lyophilized
Quantity 50 µg
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 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

AS10 1611 | anti-STN7 | serine/threonine-protein kinase STN7, chloroplastic for Arabidopsis thaliana, rabbit antibodies

AS15 3080 | anti-STN7 | serine/threonine-protein kinase STN7, chloroplastic for Chlamydomonas reinhardtii, rabbit antibodies

Secondary antibodies

Additional information An extract from STN8 mutant needs to be used in pararel to determine specific band of STN8 protein on a western blot.
application information
Recommended dilution 1 : 2000 (WB)
Expected | apparent MW

54.9 kDa or 46 kDa on 6 M urea gel

Confirmed reactivity Arabidopsis thaliana, Triticum aestivum
Predicted reactivity Oryza sativa
Not reactive in No confirmed exceptions from predicted reactivity are currently known.
Additional information

For best results with this antibody sample buffer needs to contain 6 M urea (138mM TrisHCl pH 6.8, 6M urea, 22.2% Glycerol, 4.3% SDS).

STN8 is a nuclear encoded protein which is localized in chloroplast, hence posses a chloroplastic target peptides (cTP) at the beginning of the amino acid sequence which is cut off. Accroding to TargetP (program that predicts the length of the cTP:s) the length of cTP for Stn8 is 49 amino acids. STN8 portein mobility can be also affected by urea present in the gel.

Due to a high background signal with LHCII it is adviced to cut off a membrane below 30 kDa marker.

Selected references Li et al. (2015). Effect of hydrogen sulfide on D1 protein in wheat under drought stress. Acta Physiologiae Plantarum November 2015, 37:225.
Flood et al. (2014). Natural variation in phosphorylation of photosystem II proteins in Arabidopsis thaliana: is it caused by genetic variation in the STN kinases? Philos Trans R Soc Lond B Biol Sci. 2014 Mar 3;369(1640):20130499. doi: 10.1098/rstb.2013.0499. Print 2014.
Yin et al. (2012). Photosystem II Function and Dynamics in Three Widely Used Arabidopsis thaliana Accessions. PLOS ONE, open access.

application example

western blot using anti-STN( antibodies


Thylakoids (2 or 3 µg of chlorophyll/lane) from Arabidopsis thaliana wide type (WT) and a stn8 mutant were isolated according to Sirpiö et al (2011, Methods Mol Biol. 2011; 775:19-30). Denaturated samples were separated on 12 % SDS-PAGE with 6 M urea and blotted for 1h to a PVDF membrane using a semi-dry transfer. Blots were blocked with 5% milk in TBS for 1h at room temperature (RT) with slow agitation. Blots were incubated in primary antibodies at a dilution of 1: 1 000 overnight at 4°C with slow agitation in 1 % milk/TTBS. Blots were rinsed briefly once, then washed twice for 10 min with TTBS at RT with vigorous agitation. Blots were incubated in secondary antibodies (anti-rabbit IgG horse radish peroxidase conjugated) in 1% milk/TTBS for 2 hours at RT with slow agitation, washed as above and incubated for 5 min with ECL solution according to the manufacturers’ instructions. Exposure time was 2 min. Since the STN8 antibody cross-reacts with LHCII, the lower part of the gel should be cut out before blotting.

Courtesy Virpi Paakkarinen, University of Turku, Finland


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