Assay Principle

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.

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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.

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Storage

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

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艾美捷細(xì)胞失巢凋亡檢測(cè)試劑盒提供比色和熒光格式測(cè)量錨定非依賴性生長(zhǎng)并監(jiān)測(cè)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(cè)定24個(gè)樣品?；罴?xì)胞是用MTT或Calcein AM檢測(cè)。用埃塞俄比亞同二聚體（EthD-1）檢測(cè)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(guò)MTT或鈣素AM。通過(guò)Ethidium Homodimer（EthD-1）測(cè)量失巢細(xì)胞死亡。EthD-1是一種用于測(cè)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)胞相互作用之前實(shí)際上是非熒光的。

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艾美捷細(xì)胞失巢凋亡檢測(cè)試劑盒組分：

1.抗錨固板（零件號(hào)108001）：一塊24孔Poly-Hema涂層板。

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

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

4.洗滌劑溶液（零件號(hào)108004）：一瓶–25.0 mL。

5.MTT溶液（零件號(hào)113502）：三管，每管1.0 mL。

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艾美捷細(xì)胞失巢凋亡檢測(cè)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

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下圖顯示了CytoSelect的典型結(jié)果細(xì)胞失巢凋亡檢測(cè)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：人前胸成纖維細(xì)胞BJ-TERT細(xì)胞的失活測(cè)定。BJ-TERT細(xì)胞以50000個(gè)細(xì)胞/孔接種在組織培養(yǎng)對(duì)照板或Poly-Hema涂層板中。細(xì)胞是允許培養(yǎng)24小時(shí)。通過(guò)MTT和Calcein AM測(cè)定細(xì)胞活力用EthD-1染色失巢樣細(xì)胞死亡。

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細(xì)胞失巢凋亡檢測(cè)試劑盒部分文獻(xiàn)引用：

1. Lu, S. et al. (2016). Expression of α-fetoprotein 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.