高感度HRP標識タンパク質
高感度ELISA, ウエスタンブロット, 免疫組織染色用HRP標識ビオチン結合性タンパク質
最高感度が要求されるELISAやウエスタンブロッティングでの使用を想定してデザインされた西洋ワサビパーオキシダーゼ(HRP)標識タンパク質です。HRP標識ストレプトアビジンとHRP標識ニュートラアビジンは化学発光検出, 化学蛍光検出, 発色検出法のELISAやウエスタンブロットで使用できます。安定剤入りの溶液は、4℃で1年間安定で、使い易いReady-to-Useなパッケージになっています。
特長
- 微量の目的タンパク質を高感度、高S/N比で検出
- ウエスタンブロッティングやELISAでHRPコンジュゲート使用量を削減
- 一般的なHRP用の化学発光基質、化学蛍光基質、発色基質に対応
- 安定化剤入りのReady-to-Useな溶液、4℃で1年間
Thermo Scientific High Sensitivity HRP Conjugate は目的化合物の検出, シグナル強度, シグナル/ノイズ比において従来製品を上回っています。High Sensitivity Streptavidin-HRPを用いた実験では従来品の8-10倍希釈しても、なお同様の結果が得られています。
Fig.1 Thermo Scientific High Sensitivity Streptavidin-HRP enables low-level target detection with high signal-to-noise ratios.
Recombinant GST (Cat # 20237) was serially diluted (0-10,000 pg/ml) with StartingBlock (PBS) Blocking Buffer (Cat. #37538). Each dilution (100 µl) was added to a 96-well glutathione-coated plate (Cat # 15340) in four replicates, including a negative control. The plate was incubated for 60 minutes at room temperature and washed three times with PBS Tween-20 Detergent (Cat # 28348). Biotinylated anti-GST (100 µl at 250 ng/ml; Santa Cruz) was added to all wells. The plate was incubated for 30-60 minutes at room temperature and washed three times with wash buffer. Streptavidin-HRP conjugates were diluted in blocking buffer as per manufacturer's directions. High Sensitivity Streptavidin-HRP was diluted 1:10,000 and conjugates from other suppliers were diluted to 1:1,000. The conjugate solutions (100 µl) were added to the plate and incubated for 60 minutes (light protected) at room temperature. The plate was washed five times with wash buffer and 150 µl/well of SuperSignal ELISA Pico Substrate (Cat. #34080) was added. Signal intensity was measured using a luminometer.
Fig.2 Thermo Scientific High Sensitivity HRP Conjugates provide superior performance in Western blotting.
Western blot of identical HeLa lysate samples probed with a biotinylated anti-GAPDH antibody (1 µg/ml) and then with respective streptavidin-HRP conjugates. A dilution of 1:4,000 was used for Thermo Scientific High Sensitivity Streptavidin-HRP, and a dilution of 1:1,000 was used for conjugates from the other suppliers. After washing, Thermo Scientific Pierce ECL Substrate (Cat. #32106) was added using identical conditions. Membranes were exposed to film for 5 seconds.
Fig.3 Excellent IHC staining of GAPDH and cytokeratin 18 in human colon carcinoma with Thermo Scientific High Sensitivity HRP Conjugates.
Formalin-fixed paraffin-embedded human colon carcinoma tissues were stained using Metal Enhanced DAB Substrate (Thermo Scientific Pierce Immunohistochemistry Peroxidase Detection Kit, (Cat. #36000). The tissues were incubated with either biotinylated anti-GAPDH or rabbit anti-cytokeratin 18 followed by biotinylated anti-rabbit IgG (left panels) or blocking buffer only (right panels). The sections were subsequently incubated with Thermo Scientific High Sensitivity NeutrAvidin-HRP (top panels) or Streptavidin-HRP (bottom panels). Tissues were counterstained using the Harris-modified hematoxylin solution (blue staining in all panels). GAPDH and Cytokeratin 18, stained using the High Sensitivity HRP Conjugates, appear brown (left panels) while the Negative Control panels shows no brown staining in the tissue.
| Cat # | 製 品 名 | 容 量 | 価 格 |
| 21130 | High Sensitivity Streptavidin-HRP Concentration: 1 mg/ml Also consider pre-diluted (10 µg/ml) format (Cat # 21134) | 0.5 ml | ¥34,000 |
| 21132 | High Sensitivity Streptavidin-HRP Concentration: 1 mg/ml |
5 ml | ¥171,000 |
| 31030 | High Sensitivity NeutrAvidin-HRP Concentration: 1 mg/ml |
0.5 ml | ¥29,500 |
| 31032 | High Sensitivity NeutrAvidin-HRP Concentration: 1 mg/ml |
5 ml | ¥153,000 |
