antibodies against psaa, psbe/f, peta, petc, pc, psad1 cpfbpase1  (Agrisera)

Journal: Nucleic Acids Research

Article Title: In vivo stabilization of endogenous chloroplast RNAs by customized artificial pentatricopeptide repeat proteins

doi: 10.1093/nar/gkab390

Figure Lengend Snippet: Chloroplast genome-wide analysis of RNAs associated with dPPR rbcL in vivo . ( A ) Immunoblot analysis of an immunoprecipitation of dPPR rbcL performed with HA antibodies on stroma extracts from the complemented mrl1:dPPR rbcL or Col-0 Arabidopsis plants (negative control). Pel : immunoprecipitation pellet, Sup : supernatant; IgG : Immunoglobuline G. IgGs in the experimental pellet are detected by the secondary antibody used to probe the immunoblot. A portion of the blot stained with Coomassie blue is shown to display equal loading. ( B ) RIP-seq analysis of chloroplast RNAs that associates with dPPR rbcL in vivo . The mean coverage in kiloreads per nucleotide of two replicates for the experimental dPPR rbcL immunoprecipitation and negative Col-0 control are plotted along the chloroplast genome on the same graph. Since the read coverage was too high for the chloroplast rRNA loci, these were split and displayed in a separate graph to the right. The main peaks are labeled with the names of the locus they belong to. Data for replicate experiments are shown in and the read counts are provided in . ( C ) Local enrichment (Ratio dPPR rbcL /Col-0) of rbcL RNA. The highest enrichment (more than 700-fold) is found in the 5′ UTR region where the binding site of dPPR rbcL is located. The positions of the rbcL initiation (ATG) and stop (TAG) codons are indicated with their genomic position. ( D ) Slot blot hybridization analysis of RNAs that coimmunoprecipitate with dPPR rbcL in vivo . The experimental and control immunoprecipitations were both performed with mrl1:dPPR rbcL stroma but used different antibodies: α-HA that detects dPPR rbcL or α-Myc as a negative control. Four replicate blots were hybridized with strand and gene specific oligonucleotide probes. The same blot was used for hybridization with psbT and psbE probes, psbC and psaB probes and psbA and atpF probes after stripping. The hybridization signals in the pellet and supernatant of dPPR rbcL immunoprecipitation were quantified for each gene and their ratio is displayed to the right.

Article Snippet: PsbE and PsbA (D1) antibodies were purchased from Agrisera.

Techniques: Genome Wide, In Vivo, Western Blot, Immunoprecipitation, Negative Control, Staining, Control, Labeling, Binding Assay, Dot Blot, Hybridization, Stripping Membranes

Journal: Scientific Reports

Article Title: The N-terminal sequence of the extrinsic PsbP protein modulates the redox potential of Cyt b 559 in photosystem II

doi: 10.1038/srep21490

Figure Lengend Snippet: NaCl-washed PSII membranes were cross-linked with pN15 at a molar pN15:PSII ratio of 50:1 to 200:1. For each sample, an amount of protein corresponding to 3 μg Chl was loaded onto each lane and detected with antisera against PsbE ( A ), PsbF ( B ), PsbO ( C ), and D1 ( D ). The arrow at ~11 kDa indicates the peptide of pN15 cross-linked to PsbE. The dashed arrow at ~13 kDa indicates the putative peptide of PsbE cross-linked to PsbF. The original positions of PsbE (9 kDa), PsbF (4 kDa), PsbO (33 kDa), and D1 (32 kDa) subunits are also indicated.

Article Snippet: Rabbit antibodies against PsbE and PsbF were purchased from Agrisera AB, Sweden.

Techniques:

Journal: The Journal of Biological Chemistry

Article Title: The Conserved His-144 in the PsbP Protein Is Important for the Interaction between the PsbP N-terminus and the Cyt b 559 Subunit of Photosystem II *

doi: 10.1074/jbc.M112.385286

Figure Lengend Snippet: Cross-linking of PsbP with PSII membranes using EDC and sulfo-NHS. NaCl-washed PSII membranes were incubated with 6.25 mm EDC and 5 mm sulfo-NHS in either (A) the presence or (B) the absence of WT PsbP. Cross-linked proteins corresponding to 2 μg of Chl were loaded onto each lane. The proteins were immunodetected with antisera against CP47, CP43, PsbO, D2, D1, PsbP, PsbR, and PsbE, as shown on each lane. Protein size markers are shown on the left. C, NaCl-washed PSII was treated with 6.25 mm EDC and 5 mm sulfo-NHS to activate the carboxyl groups on the PSII. After activation, the PSII was washed and incubated either in the presence (+PsbP) or in the absence (−PsbP) of PsbP. As a positive control, the sample used in A was analyzed simultaneously.

Article Snippet: Rabbit antibodies against D2, PsbR and PsbE were purchased from Agrisera.

Techniques: Incubation, Activation Assay, Positive Control