基本描述

产品名称

Annexin V-AF488

别名

重组人Annexin V/ANXA5蛋白 | 重组人膜连蛋白A5

英文别名

规格或纯度

BioReagent, 生物染色剂, 用于显微镜, 适用于免疫荧光(IF), 5 μL/test

生化机理

The placental anticoagulant protein Annexin A5 (ANXA5) is a multifunctional protein that is highly expressed on the apical surfaces of syncytiotrophoblasts, and plays an important role in haemostatic regulations, maintaining blood fluidity of the placenta. Annexin A5 (ANXA5) is a protein abundantly expressed in normal placenta where it contributes to the healthy outcome of a pregnancy. Lower ANXA5 levels have been observed in M2/ANXA5 haplotype carrying chorion. The association found between the maternal carriage of the M2/ANXA5 haplotype and an elevated risk of IUGR and/or Biotin supports the hypothesis that carrier status of this haplotype and the consequently reduced placental ANXA5 expression might be responsible, at least partially, for the onset of these gestational vascular complications. ANXA5 could be used as a biomarker for the early detection of Biotin and for the prediction of its severity. ANXA5 as an embryonic anticoagulant that appears deficient in contiguous specter of thrombophilia-related pregnancy complications culminating more frequently in miscarriage in a maternal M2 carrier background. As a potential indicator for malignancy and lymphatic metastasis, ANXA5 overexpression increases in vitro migration and invasion of Hca-P cell, promotes in vivo malignancy, LNM rate and level of Hca-P-transplanted mice. Hereditary thrombophilias can impair vascular placental functions and predispose to the birth of small-for-gestational age (SGA) babies. The placental anticoagulant protein annexin A5 (ANXA5) may contribute to this process. A functional haplotype (M2) within the ANXA5 gene is associated with fetal loss and venous thrombosis.

产品介绍

Annexin V 是一种钙离子依赖性磷脂结合蛋白，与磷脂酰丝氨酸 (PS) 有高度亲和力。当细胞发生凋亡时，膜内侧的磷脂酰丝氨酸 (PS) 外翻到膜表面，而被荧光染料标记的Annexin V结合，可通过流式细胞仪或荧光显微镜进行检测。Annexin V 搭配 PI 或 7-AAD、DAPI 等 DNA 染色液使用，可区分处于不同凋亡时期的细胞。我们提供了一系列膜联蛋白 V 和荧光染料的偶联物，包括AF405、AF488、AF647、AF680、AF750和膜联蛋白 V 偶联物以及 Annexin V APC、生物素、FITC、Pacific Blue、 PE 、PE-Cy5、PE-Cy7偶联物。这些偶联物可产生较亮信号，适用于所有可用激光，可用作独立试剂或与细胞核染色液组合成试剂盒使用。
注意事项：
1. 使用时请避光以减缓荧光淬灭；
2. PI染料具有剧毒性和致突变性，操作时需谨慎；
3. 由于膜联蛋白V（Annexin V）与磷脂酰丝氨酸（PS）的结合具有钙依赖性，因此在Annexin V实验中务必避免使用含EDTA或其他钙螯合剂的缓冲液。
使用说明：
1. 使用前需将10x 结合缓冲液（A1372288）用去离子水稀释至1x（1mL 10x缓冲液加9mL去离子水）。
2. 预冷PBS清洗细胞两次后，按照1.0-5.0x105/mL的密度用1x Annexin V结合缓冲液重悬。
3. 每100μL细胞悬液中加入5μL 荧光标记Annexin V和细胞核活性染料，例如PI (P1373641; P1372285), 7-AAD (A1372406), 或者DAPI (D1372407)等。
4. 轻柔涡旋混匀细胞，室温避光孵育10min。
5. 加入100μL结合缓冲液，1h内完成上机分析。

Annexins are a family of calcium-dependent phospholipid-binding proteins that preferentially bind phosphatidylserine (PS). Under normal physiologic conditions, PS is predominantly located in the inner leaflet of the plasma membrane. Upon initiation of apoptosis, PS loses its asymmetric distribution across the phospholipid bilayer and is translocated to the extracellular membrane leaflet marking cells as targets of phagocytosis. Once on the outer surface of the membrane, PS can be detected by fluorescently labeled Annexin V in a calcium-dependent manner.
In early-stage apoptosis, the plasma membrane excludes viability dyes such as propidium iodide (PI), 7-AAD. These cells will stain with Annexin V but not a viability dye, thus distinguishing cells in early apoptosis. However, in late stage apoptosis, the cell membrane loses integrity thereby allowing Annexin V to also access PS in the interior of the cell. A viability dye can be used to resolve these late-stage apoptotic and necrotic cells (Annexin V, viability dye-positive) from the early-stage apoptotic cells (Annexin V positive, viability dye-negative).
We offer recombinant Annexin V conjugated to a numerous fluorophores, as well as an Annexin V biotin conjugate which can be detected with fluorophore-labeled streptavidin. By binding to PS, fluorophores labeled Annexin V can be used to detect and quantify apoptotic cells via ﬂow cytometry or ﬂuorescence microscopy. The excitation and emission maxima of the Annexin V conjugates are summarized in the following table.

Cat.No.	Ex/Em (nm)	Format
rp226056	NA	Biotin
rp225999	401/422	AF405
rp226057	490/525	AF488
rp226060	650/668	AF647
rp226002	681/704	AF680
rp226003	752/776	AF750
rp226053	498/517	FITC
rp226004	410/455	Pacific Blue
rp226006	647/665	Cy5
rp226054	650/660	APC
rp226055	565/575	PE
rp226058	565/670	PE-Cy5
rp226059	565/774	PE-Cy7

Precautions

1. Please try to avoid light when using to slow down the quenching of fluorescence.

2. Propidium Iodide Solution is toxigenic and mutagenic; handle with care.

3. Due to the calcium dependence of the Annexin V:PS interaction, it is critical to avoid buffers containing EDTA or other calcium chelators during Annexin V experiments.

Instruction for use

1. Dilute 10X Binding Buffer (A1372288) to 1X using distilled water (1 mL 10X Binding Buffer + 9 mL ddH2O).

2. Wash cells twice with cold PBS and then resuspend the desired amounts of cells in Annexin V Binding Buffer at a concentration of 1.0-5.0 x 106 cells/mL.

3. Add 5 µL of Annexin V-AF488 to 100 µL of the cell suspension. Stain with a viability dye, such as PI (P1373641; P1372285), 7-AAD (A1372406), or DAPI (D1372407) dyes, if desired.

4. Gently vortex the cells and incubate for 10 min at RT (25°C) in the dark.

5. Add 100 µL of 1X Binding Buffer to each assay. Analyze by flow cytometry within 1 hr.

生物活性

Testing in progress

表达系统

E. coli

种属

人(Human)

氨基酸

2-320 aa

序列

AQVLRGTVTDFPGFDERADAETLRKAMKGLGTDEESILTLLTSRSNAQRQEISAAFKTLFGRDLLDDLKSELTGKFEKLIVALMKPSRLYDAYELKHALKGAGTNEKVLTEIIASRTPEELRAIKQVYEEEYGSSLEDDVVGDTSGYYQRMLVVLLQANRDPDAGIDEAQVEQDAQALFQAGELKWGTDEEKFITIFGTRSVSHLRKVFDKYMTISGFQIEETIDRETSGNLEQLLLAVVKSIRSIPAYLAETLYYAMKGAGTDDHTLIRVMVSRSEIDLFNIRKEFRKNFATSLYSMIKGDTSGDYKKALLLLCGEDD

蛋白标签

No tag

无载体

No

无动物源

No

偶联

AF488

Excitation(nm)

Blue Laser(488nm)

Emission(nm)

525nm

Ex/Em(nm)

Ex:490nm;Em:525nm

Accession #

P08758

来源

重组表达

储存与运输

物理形态	液体
储存缓冲液	Liquid in PBS pH 7.4, containing 1% BSA and 0.05% Proclin300.
防腐剂	0.05% Proclin300
浓度	5 μL/test
储存温度	2-8°C储存,避光,禁止冷冻
运输条件	冰袋运输
稳定性与储存	The antibody solution should be stored undiluted between 2°C and 8°C, and protected from prolonged exposure to light. Do not freeze.
分子类型	蛋白质

图片

Flow Cytometry: Annexin V-AF488 (rp226057)

Annexin V-AF488 (rp226057) - Flow Cytometry
U937 cells were either left untreated (left) or treated (right) with 4µM camptothecin (C111281) for 4 hours, then stained with Annexin V-AF488 (rp226057) and propidium iodide (PI) (P1372285) in Annexin V Binding buffer (A1372288) for 10 minutes at room temperature.

应用

应用名称	稀释比例
Flow Cytometry	5 μL/test; For flow cytometric staining, the suggested use of this reagent is 5 µL per 100,000 - million cells in a 100 µL volume of Annexin V Binding Buffer. It

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批号(Lot Number)	证书类型	货号
ZJ25F0926514	分析证书	rp226057
ZJ25F0926515	分析证书	rp226057
ZJ25F0926516	分析证书	rp226057

技术文档和文章

膜联蛋白V染色

Categories: 技术文章

探索主题:

细胞凋亡膜联蛋白V

Annexin V staining

Categories: 技术文章

探索主题:

Apoptosis Annexin V

溶液计算器

摩尔浓度计算器

计算溶液所需的质量、体积或浓度。

质量

=

浓度

x

体积

x

分子量

g/mol

稀释计算器

计算配制储备溶液所需的稀释度。

浓度1（C1）

x

体积 1 (V1)

=

浓度 2 (C2)

x

体积 2 (V2)

复溶计算器

复溶计算器允许您快速计算试剂的体积，以复溶小瓶。只需输入试剂的质量和目标浓度，计算器将确定其余部分。

体积（添加到小瓶中）

=

质量（小瓶）

÷

所需的复溶浓度

货号 (SKU)	包装规格	是否现货
rp226057-25T	25T	现货
rp226057-100T	100T	现货
rp226057-300T	300T	现货

Annexin V-AF488

库存信息

库存信息

库存信息

基本描述

储存与运输

图片

应用

质检证书（CoA,COO,BSE/TSE 和分析图谱)

技术文档和文章

溶液计算器

摩尔浓度计算器

稀释计算器

复溶计算器