Cells are cultured in poly-Hema coated plate or control plate. Cell viability is determined by MTT or

Calcein AM. Anoikis propelled cell death is measured by Ethidium Homodimer (EthD-1). EthD-1 is

an excellent marker for measuring dead cells. EthD-1 is a red fluorescent dye that can only penetrate

damaged cell membranes. EthD-1 will fluoresce with a 40-fold enhancement upon binding ssDNA,

dsDNA, RNA, oligonucleotides, ａnd triplex DNA. Background fluorescence levels are very low

because the dyes are virtually non-fluorescent before interacting with cells.

Kit Components

1. Anchorage Resistant Plate (Part No. 108001): One 24-well Poly-Hema coated plate.

2. Calcein AM (500X) (Part No. 108002): One vial – 50 µL in DMSO.

3. Ethidium Homodimer (EthD-1) (500X) (Part No. 108003): One vial – 50 µL.

4. Detergent Solution (Part No. 108004): One bottle – 25.0 mL.

5. MTT Solution (Part No. 113502): Three tubes – 1.0 mL each.

Storage

Store the Calcein AM ａnd Ethidium Homodimer at -20ºC. Store all other components at 4ºC.

艾美捷細(xì)胞失巢凋亡檢測試劑盒提供比色和熒光格式測量錨定非依賴性生長并監(jiān)測失巢細(xì)胞死亡。該套件包含足夠的試劑用于在Poly-Hema涂層的24孔板中測定24個(gè)樣品�；罴�(xì)胞是用MTT或Calcein AM檢測。用埃塞俄比亞同二聚體（EthD-1）檢測細(xì)胞死亡。細(xì)胞在聚Hema涂層板或?qū)φ瞻逯信囵B(yǎng)。通過MTT或鈣素AM。通過Ethidium Homodimer（EthD-1）測量失巢細(xì)胞死亡。EthD-1是一種用于測量死細(xì)胞的優(yōu)良標(biāo)記。EthD-1是一種紅色熒光染料，只能穿透受損的細(xì)胞膜。EthD-1在結(jié)合ssDNA時(shí)將以40倍的增強(qiáng)發(fā)出熒光，dsDNA、RNA、寡核苷酸和三鏈DNA。背景熒光水平非常低因?yàn)槿玖显谂c細(xì)胞相互作用之實(shí)際上是非熒光的。

艾美捷細(xì)胞失巢凋亡檢測試劑盒組分：

1.抗錨固板（零件號108001）：一塊24孔Poly-Hema涂層板。

2.鈣黃素AM（500X）（零件號108002）：一小瓶–50µL DMSO。

3.埃塞俄比亞同二聚體（EthD-1）（500X）（零件號108003）：一小瓶–50µL。

4.洗滌劑溶液（零件號108004）：一瓶–25.0 mL。

5.MTT溶液（零件號113502）：三管，每管1.0 mL。

艾美捷細(xì)胞失巢凋亡檢測試劑盒相關(guān)研究：

1. CBA-081: CytoSelect™ 96-Well Anoikis Assay

2. CBA-230: Cellular Senescence Detection Kit (SA-β-Gal Staining)

3. CBA-231: 96-Well Cellular Senescence Assay (SA β-Gal Activity)

4. CBA-232: Quantitative Cellular Senescence Assay (SA β-Gal)

5. CBA-240: CytoSelect™ Cell Viability ａnd Cytotoxicity Assay

下圖顯示了CytoSelect的典型結(jié)果細(xì)胞失巢凋亡檢測試劑盒。以下數(shù)據(jù)僅供參考。該數(shù)據(jù)不應(yīng)用于解釋實(shí)際后果。

圖：人胸成纖維細(xì)胞BJ-TERT細(xì)胞的失活測定。BJ-TERT細(xì)胞以50000個(gè)細(xì)胞/孔接種在組織培養(yǎng)對照板或Poly-Hema涂層板中。細(xì)胞是允許培養(yǎng)24小時(shí)。通過MTT和Calcein AM測定細(xì)胞活力用EthD-1染色失巢樣細(xì)胞死亡。

細(xì)胞失巢凋亡檢測試劑盒部分文獻(xiàn)引用：

1. Lu, S. et al. (2016). Expression of α-ferotein in gastric cancer AGS cells contributes to invasion

and metastasis by influencing anoikis sensitivity. Oncol Rep. 35:2984-2990.

2. Lee, H.W. et al. (2013). Tpl2 kinase impacts tumor growth ａnd metastasis of clear cell renal cell

carcinoma. Mol Cancer Res. 11:1375-1386.

3. Sisto, M. et al. (2009). Fibulin-6 expression ａnd anoikis in human salivary gland epithelial cells:

implications in Sjogren＇s syndrome. Int. Immunol. 21:303-311.

4. Liu, H. et al. (2008). Cysteine-rich protein 61 ａnd connective tissue growth factor induce deadhesion ａnd anoikis of retinal pericytes. Endocrinology 149:1666-1677.

來源：https://www.amyjet.com/products/CBA-080.shtml