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PsbA | D1 protein of PSII, phosphorylated

AS13 2669 | Clonality: Polyclonal | Host: Rabbit | Reactivity: [global antibody] for higher plants

PsbA | D1 protein of PSII, phosphorylated in the group Antibodies for Plant/Algal  / Global Antibodies at Agrisera AB (Antibodies for research) (AS13 2669)

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Product name, number (Agrisera, Sweden)

Data sheet Product citations Protocols Customer reviews
 

Product Information

Immunogen

KLH-conjugated synthetic peptide derived from available plant PsbA sequences with phosphorylated (T), including Arabidopsis thaliana UniProt: P83755, TAIR:AtCg00020, Oryza sativa P0C434 and other higher plant PsbA sequences

Host Rabbit
Clonality Polyclonal
Purity Affinity purified serum in PBS, pH 7.4
Format Lyophilized
Quantity 50 µg
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)
Recommended dilution 1 : 10 000 (WB)
Expected | apparent MW

38 | 28-30 kDa

Reactivity

Confirmed reactivity Arabidopsis thaliana, Chlamydomonas reinhardtii, Hordeum vulgare, Zea mays
Predicted reactivity

Dictos, Conifers, Monocts


Species of your interest not listed? Contact us
Not reactive in Cyanobacteria

Application examples

Application examples Application example

western blot using anti-phosphorylated PsbA
5 µg of total protein from 10 days old seedlings of Hordeum vulgare exposed to high light of 450 µmol to induce phosphorylation (1), or low light of 2 µmol  (2), darkness (3) for three hours were extracted with Protein Extraction Buffer PEB (AS08 300). All three samples were treated with lambda protein phosphatase (NEB) to dephosphorylate the PsbA protein (Phosphatase-treated). Samples were diluted with 1X sample buffer (NuPAGE LDS sample buffer (Invitrogen) supplemented with 50 mM DTT and heated at 70°C for 5 min and keep on ice before loading. Protein samples were separated on NuPAGE 4-12% Tris-Bis gel (Invitrogen) LDS-PAGE and blotted for 1h to 1.5h on PVDF using tank transfer. Blots were blocked immediately following transfer in 2% blocking reagent or 5% non-fat milk dissolved 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 anti-PsbA (AS05 084, first 3 and last 3 lanes on a blot) and anti-phosphorylated PsbA (AS13 2669, middle lanes) at a dilution of 1: 10 000 (in blocking reagent) for 1h at room temperature with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, and then washed 1x15 min and 3x5 min with TBS-T at room temperature with agitation. Blots were incubated in secondary antibody (Agrisera anti-rabbit IgG horse radish peroxidase conjugated, recommended secondary antibody AS09 602) diluted to 1:25 000 in blocking reagent for 1h at room temperature with agitation. The blots were washed as above. The blot was developed for 5 min with extreme femtogram range chemiluminescnce 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). Exposure time was 2 minutes. 

Additional information

This antibody is detecting phosphorylated PsbA protein.

Antibodies are purified on a non-phosphorylated peptide.

Related products

Related products

AS05 084 | Anti-PsbA | D1 protein of PSII, C-terminal, rabbit antibodies
AS05 084PRE | PsbA | D1 protein of PSII, C-terminal, pre-immune serum

AS01 016 | Anti-PsbA C-terminal, chicken antibodies
AS11 1786 | Anti-PsbA N-terminal, rabbit antibodies

AS01 016S | PsbA protein standard for quantitation and positive control

AS10 704 | Anti-PsbA | D1 protein of PSII, DE-loop, rabbit antibodies

Plant and algal protein extraction buffer

Background

Background

The psbA gene has been cloned from many species of plants, green algae, and cyanobacteria. The psbA gene is located in the chloroplast genome and encodes for the D1 protein, a core component of Photosystem II. PsbA/D1 is rapidly cycled under illumination in all oxygenic photobionts. Tracking PsbA pools using the Global PsbA antibody can show the functional content of Photosystem II in a wide range of samples.

Alternative names: 32 kDa thylakoid membrane protein, photosystem II protein D1.

Product citations

Selected references Upadhyaya and Jagadeeshwar Rao (2019). Reciprocal regulation of photosynthesis and mitochondrial respiration by TOR kinase in Chlamydomonas reinhardtii. Plant Direct Volume 3, Issue 11.
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. https://doi.org/10.3389/fpls.2017.02154.
Li et al. (2015). Effect of hydrogen sulfide on D1 protein in wheat under drought stress. Acta Physiologiae Plantarum November 2015, 37:225.

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