チラコイド膜関連タンパク質抗体

※ 本製品は研究用です。研究用以外には使用できません。

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20 µg of chloroplast fraction from (1) Arabidopsis thaliana （シロイヌナズナ）and (2) Spinacia oleracea（ホウレンソウ）were separated on 12 % SDS-PAGE and blotted 1h to PVDF. Blots were blocked with 2 % non-fat milk powder in 1xTBS-T for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 5,000 for 1h at RT 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 RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1:10,000 in for 1h at RT with agitation. The blot was washed as above and developed for 5 min with ECL according to the manufacturer's instructions.

5 µg of total protein from (1) Synechocystis sp. PCC 6803（シネコシスティス） extracted with PEB (#AS08-300), (2) Synechocystis sp. PCC 6803 grown in low iron culture, extracted with PEB, 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: 1,000 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, from Abcam) diluted to 1:20,000 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).

2 µg of total protein from (1) Arabidopsis thaliana leaf（シロイヌナズナ葉） extracted with Protein Extration Buffer, PEB (#AS08-300), (2) Hordeum vulgare leaf （オオムギ葉）extracted with PEB, (3) Chlamydomonas reinhardtii （クラミドモナス）total cell extracted with PEB, (4) Synechococcus sp. 7942（シネココッカス7942株） total cell extracted with PEB, (5) Anabaena sp.（アナベナ） total cell extracted with PEB 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).

植物組織や藻類の細胞を効率良く溶解し、総タンパク質を可溶化するための抽出用バッファーです。
詳細はこちらをご覧下さい。

光合成タンパク質関連抗体、スタンダード、二次抗体をセットにしたウエスタンブロット測定用のキットです。
詳細は「Photosynthesis Tool Kit / Educational Antibody Kit」をご覧下さい。