使用 pHrodo pH 值敏感探头染料(包括 pHrodo Red、Green 和 Deep Red 染料),可进行低信背比和高特异性细胞内化研究。这些染料可与生物分子(如抗体)偶联,且仅在酸性环境(如细胞溶酶体和内体环境)中产生高强度荧光。这可在活细胞检测中检测偶联的生物分子,而无需在实验工作流程中进行洗涤步骤或淬灭染料。
pHrodo Red、Green 和 Deep Red 染料 具有胺反应性和 pH 值敏感性的 pHrodo Red、Green 和 Deep Red 染料优化用于生成生物偶联物以用于活细胞检测中抗体内化、内吞作用和吞噬作用的研究。胺反应性 pHrodo 染料使您能够在抗体、蛋白、细胞、病毒或其他含胺生物分子上标记伯胺,以形成在酸性环境中发出荧光但在中性 pH 值环境下几乎不显示信号的偶联物。pHrodo 染料在典型细胞外环境中无荧光,使得无需在实验工作流程中进行洗涤步骤或淬灭染料。
pHrodo iFL Red 和 Green STP 酯染料 pHrodo iFL Red 和 Green STP 酯染料是原始 pHrodo Red 和 Green 染料的改进版,经过优化可比原始染料产生更多可溶性偶联物。这种增强的特性使其可用于标记在偶联过程中可能从溶液中沉淀的抗体。
pHrodo Deep Red TFP 酯 pHrodo Deep Red TFP 酯与 pHrodo iFL Red 和 Green 酯染料的不同之处在于,前者具有较低的 pKa,因此在随后内吞途径中显示出改善的信背比和荧光。我们的低背景 pHrodo Deep Red 染料使研究内化比以往更容易,结果确定性更高且优化步骤更少,因为 pHrodo Deep Red 染料仅在晚期内体和溶酶体中发荧光。
当 pH 环境酸化时,pHrodo 染料的荧光增加与细胞内囊泡酸化相关,这使得这些染料成为研究内吞作用或吞噬作用及其受环境因素、药物或病原体调节的宝贵工具。pHrodo 染料的光谱特性也使其可用于多色实验。pHrodo iFL Red、iFL Green 和 Deep Red 染料可通过大多数标准红色、绿色和深红色荧光滤光片组进行检测。pHrodo 染料偶联物经验证可用于多种应用,包括流式细胞分析、荧光显微镜检测以及高内涵筛选 (HCS)。
pHrodo Deep Red 染料具有较低 pKa 设计,在中性条件下无信号,仅在酸性环境中发荧光。这种独特的特性可在研究抗体内化和内吞/吞噬途径时实现快速检测开发,并提高结果的确定性。pHrodo Deep Red 染料有助于更好地鉴别来自细胞外的内化物质,而 pHrodo iFL Red 和 Green 染料则会在早期内体中发出荧光。
规格
化学反应性胺
描述pHrodo Deep Red TFP 酯
激发/发射640/655
标签或染料pHrodo™ 深红色
分子量∼1300
数量3 x 100μg
反应一部分TFP 酯
运输条件室温
颜色深红色
标签类型pH 值敏感的染料
产品线pHrodo™
Unit SizeEach
内容与储存
在冷冻冰箱(-5°C 至 -30°C)中避光储存。
常见问题解答 (FAQ)
May I store pHrodo Deep Red TFP Ester (Cat. Nos. P35358, P35259) labeled antibodies or other proteins in solutions containing sodium azide?
We do not recommend storing antibodies or proteins labeled with pHrodo Deep Red TFP Ester (Cat. Nos. P35358, P35359) in solutions containing sodium azide, as pHrodo Deep Red is not stable in the presence of sodium azide. Consider using an alternative, non-azide-based anti-microbial.
After labeling with pHrodo Deep Red TFP Ester (Cat. Nos. P35358, P35259), may I store the dye-protein conjugated in other buffers?
Yes, after labeling with pHrodo Deep Red TFP Ester (Cat. Nos. P35358, P35259) you can store your dye-conjugated protein in other buffers based on what is ideal for maintaining the stability of your protein. We do not recommend storing pHrodo Deep Red-labeled proteins in solutions containing sodium azide.
After labeling with pHrodo Deep Red TFP Ester (Cat. Nos. P35358, P35259), do I have to store my protein at 2–8 degrees C or can I store it at other temperatures?
Yes, after labeling with pHrodo Deep Red TFP Ester (Cat. Nos. P35358, P35259) you can store your dye-conjugated protein at other temperatures based on what is ideal for maintaining the stability of your protein.
Does my protein have to be at an exact concentration of 2.2 mg/mL as noted in the labeling protocol for pHrodo Deep Red TFP Ester (Cat. Nos. P35358, P35359)?
The final protein concentration of ~2 mg/mL for the labeling reaction of pHrodo Deep Red TFP Ester (Cat. Nos. P35358, P35359) affects good reaction kinetics and good labeling with the recommended range of the dye-to-protein molar ratio (MR) from 5-20 moles of dye per mole of protein. You may use a protein concentration >2 mg/mL assuming the solution does not become more viscous, or the protein becomes more aggregated and, optimize the MR.