D1 Protein of PSII, C-terminal (PsbA) antibody

Antigen: D1 Protein of PSII, C-terminal (PsbA)

Clonality: Polyclonal

Host: Chicken

Reactivity: Seaweed (Alaria esculenta), Amphidinium carterae (A. carterae), Arabidopsis thaliana, Chlamydomonas reinhardtii (C. reinhardtii), Chlamydomonas raudensis, Cyanophora, Gonyaulax polyedra, Fucus vesiculosus, Barley (Hordeum vulgare), Lobaria pulmonaria, Petunia, Pinus sylvestris, Synechococcus PCC 7942, Ulva

Application: Western Blotting (WB)

Catalog no.: ABIN349641

Additional Information

Immunogen: KLH-conjugated synthetic peptide derived from available plant,algal andcyanobacterial PsbA sequences, including Arabidopsis thaliana AtCg00020 ,Oryza sativa P0C434, Populus alba Q14FH6, Synechocystis sp. P14660 andmany others0

Format: Liquid

Description: The PsbA (D1) protein of Photosystem II is rapidly cycled under illumination in alloxygenic photobionts. Disruption of PsbA cycling or losses of PsbA pools arecentral to photoinhibition of photosynthesis in cyanobacteria, algae and plantsunder a wide range of conditions including excess light, low temperature and UVexposure. Tracking PsbA pools using the Global PsbA antibody can show thefunctional content of Photosystem II in a wide range of samples.

Specificity: Predicted reactivity: dicots including legumes, monocots, conifers, brown algae, red algae,cryptomonads, stramenopiles, euglenoids, xantophytes, prochlorophytes.

Application Details

Application Notes: Recommended Dilution 1:4000 - 1: 8000 on 5 µg of total protein, detected with standard ECL (WB). Additional Information: A number of degradation products may be observed when using anti-PsbAantibodies, including products having apparent molecular weights of 24kDa and16kDa. D1 degradation is a complex set of events and the products observed canbe influenced by both the extraction procedure and the physiology of the cellsprior to harvest. Third, cross-linking may occur between D1 and cytochrome b559,shifting the protein higher in the gel. In cyanobacteria (PCC7942), three differentbands were competed out by preincubating the antibody with the PsbA freepeptide, indicating that all bands are indeed PsbA and its precursors orbreakdown product.

Buffer: total IgY in PBS pH 8.0+ 0.02% sodium azide

Storage: store at 4°C, make aliquots to avoid working with a stock. Please, Remember tospin tubes briefly prior to opening them to avoid any losses that might occur fromliquid material adhering to the cap or sides of the tubes.

Research Area: Plant/Algal Physiology

Restrictions: For Research Use only

Images

2ug of total protein from (1) Arabidopsis thaliana leaf extracted with PEB , (2) Horderum vulgare leaf extracted with PEB , (3) Chlamydomonasreinhardtii total cell extracted with PEB , (4) Synechococcus sp. 7942 totalcell extracted with PEB , (5) Anabaena sp. total cell extracted with PEB were separated on 4-12% NuPage (Invitrogen) LDS-PAGE and blotted1h to PVDF. Blots were blocked immediately following transfer in 2% ECL Advanceblocking reagent (GE Healthcare) in 20 mM Tris, 137 mM sodium chloride pH 7.6 with0.1% (v/v) Tween-20 (TBS-T) for 1h at room temperature with agitation. Blots wereincubated in the primary antibody at a dilution of 1: 50 000 for 1h at room temperaturewith agitation. The antibody solution was decanted and the blot was rinsed brieflytwice, then washed once for 15 min and 3 times for 5 min in TBS-T at roomtemperature with agitation. Blots were incubated in secondary antibody (anti-hen IgYhorse radish peroxidase conjugated, from Abcam) diluted to 1:50 000 in 2% ECLAdvance blocking solution for 1h at room temperature with agitation. The blots werewashed as above and developed for 5 min with ECL Advance detection reagentaccording the manufacturers instructions. Images of the blots were obtained using aCCD imager (FluorSMax, Bio-Rad) and Quantity One software (Bio-Rad).

Publications

Publications: Wang, Jensen, Huojrup et al.: "Synthesis and degradation of dinoflagellate plastid-encoded psbA proteins are light-regulated, not circadian-regulated." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 102, Issue 8, pp. 2844-9, 2005 (PubMed).

Sicora, Appleton, Brown et al.: "Cyanobacterial psbA families in Anabaena and Synechocystis encode trace, constitutive and UVB-induced D1 isoforms." in: Biochimica et biophysica acta, Vol. 1757, Issue 1, pp. 47-56, 2006 (PubMed).

Titiz, Tambasco-Studart, Warzych et al.: "PDX1 is essential for vitamin B6 biosynthesis, development and stress tolerance in Arabidopsis." in: The Plant journal : for cell and molecular biology, Vol. 48, Issue 6, pp. 933-46, 2007 (PubMed).