BCECF试剂货号:B031 CAS号:85138-49-4

BCECF试剂货号:B031
2′,7′-Bis(carboxyethyl)-4 or 5-carboxyfluorescein
CAS号:85138-49-4
商品信息
储存条件:室温
运输条件:室温
SDS 
选择规格:下载
5mg
期货 
 
荧光探针检测方案
性质
注意事项
溶解例
参考文献
规格性状

性质

  5-羧基荧光素是一种通过将染料掺入细胞来研究细胞内pH变化的方法,但是当用于pKa = 7.0的细胞(如淋巴细胞)中时,染料会从细胞中洗脱出来,很难快速测定。 BCECF引入了两个羧乙基基团以减少从细胞中的洗脱。 适用于研究细胞内pH变化(λex= 490 nm,λem= 526 nm)。

注意事项

  ・如果将本产品以粉末形式取出并使用,由于其性质,静电等因素,它可能会粘附在容器内部,从而难以完全取出。

・对于粘附在容器上且无法取出的粉末,请在使用前将要使用的溶剂倒入容器中并溶解。

溶解例

2 mg / ml(甲醇)

参考文献

1) R. A. Steinhardt and D. Mazia, “Development of K+-conductance and Membrane Potentials in Unfertilized Sea Urchin Eggs After Exposure to NH4OH”, Nature1973241, 400.
2) T. J. Rink, R. Y. Tsien and T. Pozzan, “Cytoplasmic pH and Free Mg2+ in Lymphocytes”, J. Cell Biol.198295, 189.
3) A. M. Paradiso, R. Y. Tsien and T. E. Machen, “Na+ -H+ Exchange in Gastric Glands as Measured with a Cytoplasmic-trapped, Fluorescent pH Indicator”, Proc. Natl. Acad. Sci. USA198481, 7436.
4) S. Grinstein, B. Elder and W. Furuya, “Phorbol Ester-induces Changes of Cytoplasmic pH in Neutrophils: Role of Exocytosis in Na+ – H+ Exchange”, Am. J. Physiol.1985248, C379.
5) G. B. Zavoico, E. J. Cragoe and M. B. Feinstein, “Regulation of Intracellular pH in Human Platelets”, J. Biol. Chem.1986261(28), 13160.
6) G. R. Bright, W. Fisher, J. Rogowska and L. Taylor, “Fluorescence Ratio Imaging Microscopy: Temporal and Spatial Measurements of Cytoplasmic pH”, J. Cell Biol.1987104, 1019.
7) C. Aalkjaer and E. J. Gragoe Jr, “Intracellular pH Regulation in Resting and Contracting Segments of Rat Mesenteric Resistance Vessels”, J. Physiol.1988402, 391.
8) K. Tsujimoto, M. Semadeni, M. Huflejt and L. Packer, “Intracellular pH of Halobacteria Can Be Determined by the Fluorescent Dye 2′,7′-bis(carboxyethyl)-5(6)-carboxyfluorescein”, Biochem. Biophys. Res. Commun.1988155, 123.
9) M. A. Kolber, R. R. Quinones, R. E. Gress and P. A. Henkart, “Measurament of Cytotoxicity by Target Cell Release and Retention of the Fluorescent Dye Bis-carboxyethyl-carboxyfluorescein(BCECF)”, J. Immunol. Methods, 1988108, 255.
10) H. Harada, Y. Kanai, M. Anzai and Y. Suketa, “cAMP Activates Cl/HCO3 Exchange for Regulation of Intracellular pH in Renal Epithelial Cells”, Biochim. Biophys. Acta, 1991, 1092, 404.
11) C. C. Freudenrich, E. Murphy, L. A. Levy, R. E. London and M. Lieberman, “Intracellular pH Modulates Cytosolic Free Magnesium in Cultured Chicken Heart Cells”, Am. J. Physiol., 1992, 262(4), C1024.
12) K. Khodakhah and D. Ogden, “Functional Heterogeneity of Calcium Release by Inositol Triphosphate in Single Purkinje Neurones, Cultured Cerebellar Astorocytes, and Peripheral Tissues”, Proc. Natl. Acad. Sci. USA, 1993, 90, 4976.
13) G. Boyarsky, C. Hanssen and L. A. Clyne, “Superiority of in vitro Over in vivo Calibrations of BCECF in Vascular Smooth Muscle Cells”, FASEB J., 1996, 10, 1205.
14) S. A. Weston and C. R. Parish, “New Fluorescent Dyes for Lymphocyte Migration Studies Analysis by Flow Cytometry and Fluorescent Microscopy”, J. Immunol. Methods, 1990, 133, 87.
15) L. S. D. Clerck, C. H. Bridts, A. M. Mertens, M. M. Moens and W. J. Stevens, “Use of Fluorescent Dyes in the Determination of Adherence of Human Leucocytes to Endothelial Cells and the Effects of Fluorochromes on Cellular Function”, J. Immunol. Methods, 1994, 172, 115.

规格性状

规格性状:

该产品为橙色至红棕色粉末,可溶于甲醇。

纯度(HPLC):85.0%以上

溶于甲醇:测试合格

荧光光谱:测试合格

红外光谱:测试合格

NMR光谱:测试一致

处理条件:

1.储存方法:避光

关联产品

Fluo 4-AM special packaging试剂
1-[2-Amino-5-(2,7-difluoro-6-acetoxymethoxy-3-oxo-9-xanthenyl)phenoxy]-2-(2-amino-5-methylphenoxy)ethane-N,N,N’,N’-tetraacetic acid, tetra(acetoxymethyl)ester
Fluo 4-AM试剂
1-[2-Amino-5-(2,7-difluoro-6-acetoxymethoxy-3-oxo-9-xanthenyl)phenoxy]-2-(2-amino-5-methylphenoxy)ethane-N,N,N’,N’-tetraacetic acid, tetra(acetoxymethyl)ester
Rhod 2-AM试剂
1-[2-Amino-5-(3-dimethylamino-6-dimethylammonio-9-xanthenyl)phenoxy]-2-(2-amino-5-methylphenoxy)ethane-N,N,N’,N’-tetraacetic acid, tetraacetoxymethyl ester, chloride
Fluo 3-AM试剂
Fluo3-AM1-[2-Amino-5-(2,7-dichloro-6-acetoxymethoxy-3-oxo-9-xanthenyl)phenoxy]-2-(2-amino-5-methylphenoxy)ethane-N,N,N’,N’-tetraacetic acid, tetra(acetoxymethyl)ester
Fura2-AM试剂
1-[6-Amino-2-(5-carboxy-2-oxazolyl)-5-benzofuranylo

BCECF-AM试剂货号:B262 3′-O-Acetyl-2′,7′-bis(carboxyethyl)-4 or 5-carboxyfluorescein, diacetoxymethyl ester CAS号:117464-70-7

BCECF-AM试剂货号:B262
3′-O-Acetyl-2′,7′-bis(carboxyethyl)-4 or 5-carboxyfluorescein, diacetoxymethyl ester
CAS号:117464-70-7
商品信息
储存条件:-20度保存,避光
运输条件:室温
分子式:

C35H28O15

分子量:

688.59

特点:

 

● 水溶性好,可进入细胞膜

● 荧光强度高

下载说明书
SDS下载
选择规格:
1mg
现货
荧光探针检测方案
活动进行中
规格性状
产品概述
产品特性
操作说明
文献

活动进行中

订购满5000元,200元礼品等你拿

凑单关联产品TOP5

NO.1.    Annexin V, FITC Apoptosis Detection Kit    细胞凋亡检测

NO.2.    Cell Counting Kit-8    细胞增殖毒性检测

NO.3.    Cytotoxicity LDH Assay Kit-WST    乳酸脱氢酶(LDH)检测

NO.4.    Cell Viability Assay Kit    细胞增殖/毒性检测

NO.5.    Fluo 4-AM special packaging    细胞内钙离子检测

 

规格性状

规格

 

特性:该产物为浅橙色至浅橙色棕色粉末或固体,可溶于DMSO和乙腈。

纯度(HPLC):90%以上

荧光光谱:试验成功

NMR光谱:试验成功

产品概述

BCECF被广泛地用于细胞内pH的测定。Tsien博士和其他人通过加入2个额外的羧化物基团改进了这种羧基荧光素。加入的两个羧基使得它能更好地被固定在细胞内。BCECF具有很好的水溶性,是因为它在中性pH时带有4-5个负电荷。但是BCECF却很难穿过细胞膜进入到细胞内。它的pKa值为6.97,高于其它的羧基荧光素。BCECF在激发光谱439nm处有等吸收点,因此它能和Fura 2一样可用比率法测定。505nm和439nm通常是非常有用的用于比率测量的波长,一般会将490nm和450nm的滤光片加在激发光源的前面。而530nm的滤光片则是用来查看它的荧光信号的。这里要注意的是激发光谱与吸收光谱有一些细微的区别。BCECF-AM是乙酰甲酯化的BCECF。它能够轻易地进入到细胞内。就和其他的乙酰甲酯类一样,BCECF-AM只需要通过孵育就能进入细胞。BCECF-AM对潮湿非常敏感,所以要小心保存。如果DMSO储存液的颜色从浅黄色变为深橙色就代表了AM的分解。所以通过对颜色变化的观测,能判断AM酯的水解情况。

B262-1.gif

产品特性

BCECF-AM是一种对细胞内pH敏感的新型荧光探针,分别用490nm和440nm波长激发BCECF所得到的发射荧光之比与pH有很好的线性关系。BCECF-AM是乙酰甲酯化的BCECF,BCECF-AM是一种可以穿透细胞膜的荧光染料,BCECF-AM没有荧光,进入细胞后被细胞内的酯酶水解成BCECF,从而被留在细胞内。BCECF在适当的pH值情况下可以被激发形成绿色荧光。BCECF-AM不仅被广泛用于哺乳动物细胞的研究,也有报道用于动物组织、植物细胞、细菌和酵母等的细胞内pH水平检测。在有细胞内pH变化的细胞毒性、细胞凋亡、细胞粘附、药物抵抗、细胞趋化等过程中BCECF AM被广泛应用。BCECF-AM(pH荧光探针)需用无水DMSO(anhydrous DMSO)配制。

BCECF-AM(x600)-2.jpg

操作说明

试剂 (溶解方法):   

1.45 mM的BCECF-AM/DMSO溶液(将1 mg 的BCECF-AM溶于1mlDMSO)HEPES缓冲液(20mM HEPES,153mM NaCl,5mM KCl,5mM glucose,pH7.4)

操作:

1.用HEPES制备细胞悬液,细胞浓度为4×107个/ml。

2.将1.45 mM的BCECF-AM/DMSO溶液加入细胞悬液中 (细胞悬液的1/300体积),BCECF-AM终浓度为3mM。

3.37℃培养30min。

4.用HEPES缓冲液清洗细胞3次,制成3×106个/ml的细胞悬液。

5.使用荧光显微镜或带有图像分析系统的激光共聚焦显微镜检测细胞的荧光强度。

*标记的条件因细胞种类而异,在每次实验前,请先确定最佳条件。以上方法仅供参考。

B262-3.png

文献

1.R.A.Steinhardt, et al.,Development of K+-conductance and Membrane Potentials in Unfertilized Sea Urchin Eggs After Exposure to NH4OH. Nature.1973;241:400-401.

2.T.J.Rink, et al.,Cytoplasmic pH and Free Mg2+ in Lymphocytes.J Cell Biol.1982;95:189-196.

3.A.M.Paradiso,et al.,Na+ -H+ Exchange in Gastric Glands as Measured with a Cytoplasmic-trapped, Fluorescent pH Indicator. PNAS.1984;81:7436-7440.

4.S.Grinstein,et al.,Phorbol Ester-induces Changes of Cytoplasmic pH in Neutrophils:Role of Exocytosis in Na+ – H+ Exchange.Am J Physiol.1985;248:C379-C386.

5.G.B.Zavoico, et al.,Regulation of intracellular pH in human platelets.Effects of thrombin,A23187,and ionomycin and evidence for activation of Na+/H+ exchange and its inhibition by amiloride analogs.J Biol Chem.1986;261:13160-13167.

6.G.R.Bright,et al.,Fluorescence Ratio Imaging Microscopy: Temporal and Spatial Measurements of Cytoplasmic pH.J Cell Biol.1987;104:1019-1033.

7.C.Aalkjaer,et al.,Intracellular pH Regulation in Resting and Contracting Segments of Rat Mesenteric Resistance Vessels. J Physiol.1988;402:391-410.

8.K.Tsujimoto,et al.,Intracellular pH of Halobacteria Can Be Determined by the Fluorescent Dye 2 E 7 Ebis(carboxyethyl)-5(6)-carboxyfluorescein.Biochem Biophys Res Commun.1988;155:123-129.

9.M.A.Kolber,et al.,Measurament of Cytotoxicity by Target Cell Release and Retention of the Fluorescent Dye Bis-carboxyethylcarboxyfluorescein(BCECF).J Immunol Methods.1988;108:255-264.

10.H.Harada,et al.,cAMP Activates Cl-/HCO3 – Exchange for  Regulation of Intracellular pH in Renal Epithelial Cells. Biochim Biophys  Acta. 1991;1092:404-407.11.C.C. Freudenrich, et al.,Intracellular pH Modulates  Cytosolic Free Magnesium in Cultured Chicken Heart Cells.Am J Physiol.1992;262:C1024-C1030.

12.K.Khodakhah,et al.,Functional Heterogeneity of Calcium  Release by Inositol Triphosphate in Single Purkinje Neurones, Cultured  Cerebellar Astorocytes, and Peripheral Tissues. PNAS. 1993;90:4976-4980.

13.G.Boyarsky,et al.,Superiority of in vitro Over in vivo Calibrations of BCECF in Vascular Smooth Muscle Cells. FASEB J.1996;10:1205-1212.

14.S.A. Weston,et al., New Fluorescent Dyes for Lymphocyte  Migration Studies Analysis by Flow Cytometry and Fluorescent Microscopy.J Immunol Methods.1990;133:87-97.

15.L.S.De Clerck,et al.,Use of Fluorescent Dyes in the Determination of Adherence of Human Leucocytes to Endothelial Cells and the Effects of Fluorochromes on Cellular Function.J Immunol Methods.1994;172:115-124.