BioVector? Mito-R-GECO1 線粒體紅色熒光鈣離子指示質粒 / Mito-R-GECO1 Mitochondrial Red Calcium Indicator Plasmid

通用定義 / General Definition:

BioVector? Mito-R-GECO1 是一種專用于監(jiān)測線粒體內鈣離子 ($Ca^{2+}$) 動態(tài)的基因編碼鈣指示劑 (GECI)。該質粒表達一種紅色熒光蛋白（基于 R-GECO1），并通過在 $N$ 端融合線粒體靶向序列（通常來源于 $Cytochrome$$c$$oxidase$$subunit$$VIII$, $COX8$），將指示劑精確鎖定在線粒體基質中。當線粒體內 $Ca^{2+}$ 濃度升高時，其紅色熒光強度會隨之增加，是研究線粒體代謝、凋亡及細胞信號轉導的強有力工具。

BioVector? Mito-R-GECO1 is a genetically encoded calcium indicator (GECI) specifically designed for monitoring mitochondrial calcium ($Ca^{2+}$) dynamics. The plasmid expresses a red fluorescent protein (based on R-GECO1) fused to a mitochondrial targeting sequence (typically from $Cytochrome$$c$$oxidase$$subunit$$VIII$, $COX8$). This ensures the indicator is precisely localized within the mitochondrial matrix. Upon an increase in mitochondrial $Ca^{2+}$ concentration, the red fluorescence intensity increases, making it a powerful tool for studying mitochondrial metabolism, apoptosis, and cellular signaling.

BioVector? Mito-R-GECO1 技術說明書 (Plasmid Datasheet)

中文版說明書

1. 產品基本信息

• 產品名稱： BioVector? Mito-R-GECO1 鈣指示質粒

• 指示劑類型： 強度型紅色熒光鈣指示劑 (R-GECO1)

• 靶向定位： 線粒體基質 (Mitochondrial Matrix)

• 啟動子： BioVector? CMV 強啟動子

• 抗性基因： * 大腸桿菌： kanamycin (卡那霉素) 或 Ampicillin (氨芐青霉素)

  • 真核篩選： Neomycin (G418) 或 無（取決于骨架）

• 激發(fā)/發(fā)射波長：$Ex/Em = 561 / 589$ nm

2. 培養(yǎng)與擴增條件

• 宿主菌： BioVector? DH5$\alpha$ 或 TOP10。

• 培養(yǎng)基： BioVector? LB 培養(yǎng)基。

• 抗生素： 根據載體要求添加 (如 50 $\mu$g/mL 卡那霉素)。

• 培養(yǎng)環(huán)境： 37°C 恒溫振蕩培養(yǎng)。

3. 載體應用

• 線粒體功能分析： 實時監(jiān)測刺激條件下線粒體對 $Ca^{2+}$ 的吸收與釋放。

• 多色成像： 與綠色熒光指示劑（如 GCaMP）配合，同時觀察細胞質和線粒體 $Ca^{2+}$。

• 藥物篩選： 評估藥物對線粒體鈣穩(wěn)態(tài)的影響。

4. 注意事項

• 定位驗證： 轉染后建議使用 BioVector? MitoTracker 對細胞進行共染，驗證 R-GECO1 的線粒體定位準確性。

• 光漂白： 紅色熒光蛋白雖較穩(wěn)定，但長時間高強度激發(fā)仍可能發(fā)生光漂白，建議優(yōu)化成像參數。

English Datasheet

1. General Product Information

• Product Name: BioVector? Mito-R-GECO1 Calcium Indicator Plasmid

• Indicator Type: Intensity-based Red Fluorescent Calcium Indicator (R-GECO1)

• Localization: Mitochondrial Matrix

• Promoter: BioVector? CMV constitutive promoter

• Selection Markers: * E. coli: Kanamycin or Ampicillin

  • Eukaryotic: Neomycin (G418) or None (depending on backbone)

• Excitation/Emission:$Ex/Em = 561 / 589$ nm

2. Culture and Amplification Conditions

• Host Strains: BioVector? DH5$\alpha$ or TOP10.

• Culture Medium: BioVector? LB Medium.

• Antibiotic Supplement: As per vector requirement (e.g., 50 $\mu$g/mL Kanamycin).

• Incubation: 37°C with constant shaking.

3. Applications

• Mitochondrial Function Assays: Real-time monitoring of mitochondrial $Ca^{2+}$ uptake and release under various stimuli.

• Multi-color Imaging: Co-imaging with green indicators (e.g., GCaMP) to observe cytosolic and mitochondrial $Ca^{2+}$ simultaneously.

• Drug Discovery: Evaluating the effects of compounds on mitochondrial calcium homeostasis.

4. Key Usage Notes

• Localization Verification: Post-transfection, it is recommended to co-stain with BioVector? MitoTracker to confirm the accurate mitochondrial localization of the R-GECO1 signal.

• Photobleaching: While R-GECO1 is relatively stable, prolonged exposure to high-intensity excitation light can cause photobleaching; optimization of imaging frequency and intensity is advised.

Note: For highest imaging quality, use BioVector? phenol red-free medium during live-cell imaging to reduce background fluorescence. Always ensure plasmids are extracted using BioVector? endotoxin-free kits for sensitive primary cell transfections.

Supplier生產廠家: BioVector NTCC典型培養(yǎng)物保藏中心

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