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  3. Hoechst 33258 trihydrochloride

Hoechst 33258 trihydrochloride  (Synonyms: bisBenzimide H 33258 trihydrochloride; H 33258 trihydrochloride)

目錄號(hào): HY-15558A 純度: 98.87%
COA 產(chǎn)品使用指南

Hoechst 33258 trihydrochloride 是 Hoechst 系列的標(biāo)記染料。Hoechst 系列是活細(xì)胞核標(biāo)記染料。Hoechst 通過(guò)結(jié)合 DNA 雙鏈中的小溝而結(jié)合核酸。Hoechst 傾向于結(jié)合富含 A/T 的 DNA 鏈。同時(shí),富含 A/T 的雙鏈 DNA 使熒光強(qiáng)度顯著增強(qiáng)。Hoechst 可穿過(guò)細(xì)胞膜,結(jié)合活細(xì)胞或固定細(xì)胞。Hoechst 染料的熒光強(qiáng)度隨著溶液 pH 升高而增強(qiáng)。

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Hoechst 33258 trihydrochloride Chemical Structure

Hoechst 33258 trihydrochloride Chemical Structure

CAS No. : 23491-45-4

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規(guī)格 價(jià)格 是否有貨 數(shù)量
10 mM * 1 mL in DMSO ¥820
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25 mg ¥435
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50 mg ¥623
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100 mg ¥1084
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Other Forms of Hoechst 33258 trihydrochloride:

  • 生物活性

  • 實(shí)驗(yàn)參考方法

  • 純度 & 產(chǎn)品資料

  • 參考文獻(xiàn)

生物活性

Hoechst 33258 trihydrochloride is a marker dye in Hoechst series. Hoechst is A live nuclear marker dye. Hoechst binds to the grooves in the DNA double strand, which tends to be A/ T-rich DNA strand. Although it binds to all nucleic acids, the A/ T-rich double strand DNA significantly enhances fluorescence intensity Therefore,Hoechst dye can be used for living cell labeling. The fluorescence intensity of Hoechst dye increases with the increase of pH of solution[1].

IC50 & Target

IC50: 51.31±4.56 μM (HeLa cell), 32.43±3.27 μM (HL60 cell), 15.42±2.16 μM (U937 cell)[1]

細(xì)胞效力
(Cellular Effect)
Cell Line Type Value Description References
DU-145 IC50
> 10 μM
Compound: Hoechst 33258
Cytotoxicity against human DU145 cells after 48 hrs by SRB assay
Cytotoxicity against human DU145 cells after 48 hrs by SRB assay
[PMID: 28513176]
HeLa IC50
23.52 μM
Compound: Hoechst 33258
Antiproliferative activity against human HeLa cells after 48 hrs by MTT assay
Antiproliferative activity against human HeLa cells after 48 hrs by MTT assay
[PMID: 30679134]
KG-1 IC50
42 μM
Compound: Hoechst 33258
Antiproliferative activity against human KG-1 cells assessed as inhibition of cell growth measured after 48 hrs by MTT assay
Antiproliferative activity against human KG-1 cells assessed as inhibition of cell growth measured after 48 hrs by MTT assay
[PMID: 35152694]
MCF7 IC50
48.1 μM
Compound: Hoechst 33258
Antiproliferative activity against human MCF7 cells after 48 hrs by MTT assay
Antiproliferative activity against human MCF7 cells after 48 hrs by MTT assay
[PMID: 30679134]
體外研究
(In Vitro)

Hoechst 工作液的制備
1.1 儲(chǔ)備液的制備
取 10 mg 溶于 5 mL DMSO
注意:建議儲(chǔ)備液在 4℃ 或 -20℃ 避光保存,避免反復(fù)凍融。
1.2 Hoechst 工作液的制備
稀釋儲(chǔ)備液在無(wú)血清細(xì)胞培養(yǎng)基或 PBS 中溶解,得到終濃度為 10 μg/mL 的 Hoechst工作液。
注意:請(qǐng)根據(jù)實(shí)際情況調(diào)整 Hoechst 工作液的濃度。
1.細(xì)胞染色
2.1 懸浮細(xì)胞 (6 孔板)
a.在 4℃ 下以 1000 g 離心 3-5 分鐘,然后棄去上清液。PBS 洗滌 2 次,每次 5 分鐘。細(xì)胞密度為 1×106/mL。
b.加入 1 mL 工作液,室溫孵育 3-10 分鐘。
c. 4℃、400 g 離心 3-4 分鐘,棄上清。
d.用 PBS 洗滌 2 次,每次 5 分鐘。
e.用無(wú)血清細(xì)胞培養(yǎng)基或 PBS 重懸細(xì)胞。熒光顯微鏡或流式細(xì)胞術(shù)觀察。
2.2 貼壁細(xì)胞
a.在無(wú)菌蓋玻片上培養(yǎng)貼壁細(xì)胞。
b.從培養(yǎng)基上取下蓋玻片并吸出多余的培養(yǎng)基。
c.加入 100 μL 工作液,輕搖使之完全覆蓋細(xì)胞,室溫孵育 3-10 分鐘。
d.用培養(yǎng)基洗滌 2 次,每次 5 分鐘。熒光顯微鏡或流式細(xì)胞儀觀察。

注意事項(xiàng)
1、請(qǐng)根據(jù)實(shí)際情況調(diào)整 Hoechst 工作液的濃度。
2. 本品供 R&D 使用僅供使用,不得用于藥物、家庭或其他用途。
3. 為了您的安全和健康,請(qǐng)穿著實(shí)驗(yàn)服和一次性手套進(jìn)行操作。

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

分子量

533.88

Formula

C25H27Cl3N6O

CAS 號(hào)
性狀

固體

顏色

Yellow to green

Emission (Em)

451

Excitation (Ex)

356

運(yùn)輸條件

Room temperature in continental US; may vary elsewhere.

儲(chǔ)存方式

4°C, sealed storage, away from moisture and light

*In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light)

溶解性數(shù)據(jù)
細(xì)胞實(shí)驗(yàn): 

DMSO 中的溶解度 : 100 mg/mL (187.31 mM; 超聲助溶; 吸濕的 DMSO 對(duì)產(chǎn)品的溶解度有顯著影響,請(qǐng)使用新開(kāi)封的 DMSO)

H2O 中的溶解度 : 12.5 mg/mL (23.41 mM; 超聲助溶 (<60°C))

配制儲(chǔ)備液
濃度 溶劑體積 質(zhì)量 1 mg 5 mg 10 mg
1 mM 1.8731 mL 9.3654 mL 18.7308 mL
5 mM 0.3746 mL 1.8731 mL 3.7462 mL
查看完整儲(chǔ)備液配制表

* 請(qǐng)根據(jù)產(chǎn)品在不同溶劑中的溶解度選擇合適的溶劑配制儲(chǔ)備液;一旦配成溶液,請(qǐng)分裝保存,避免反復(fù)凍融造成的產(chǎn)品失效。
儲(chǔ)備液的保存方式和期限:-80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light)。-80°C儲(chǔ)存時(shí),請(qǐng)?jiān)?個(gè)月內(nèi)使用,-20°C儲(chǔ)存時(shí),請(qǐng)?jiān)?個(gè)月內(nèi)使用。

* 備注:如您選擇水作為儲(chǔ)備液,請(qǐng)稀釋至工作液后,再用 0.22 μm 的濾膜過(guò)濾除菌后使用。

  • 摩爾計(jì)算器

  • 稀釋計(jì)算器

Mass (g) = Concentration (mol/L) × Volume (L) × Molecular Weight (g/mol)

質(zhì)量
=
濃度
×
體積
×
分子量 *

Concentration (start) × Volume (start) = Concentration (final) × Volume (final)

This equation is commonly abbreviated as: C1V1 = C2V2

濃度 (start)

C1

×
體積 (start)

V1

=
濃度 (final)

C2

×
體積 (final)

V2

動(dòng)物實(shí)驗(yàn):

以下溶解方案,請(qǐng)直接配制工作液。建議現(xiàn)用現(xiàn)配,在短期內(nèi)盡快用完。 以下溶劑前顯示的百分比是指該溶劑在您配制終溶液中的體積占比; 如在配制過(guò)程中出現(xiàn)沉淀、析出現(xiàn)象,可以通過(guò)加熱和/或超聲的方式助溶。

  • 方案 一

    請(qǐng)依序添加每種溶劑: PBS

    Solubility: 9.09 mg/mL (17.03 mM); 澄清溶液; 超聲助溶 (<60°C)

動(dòng)物溶解方案計(jì)算器
請(qǐng)輸入動(dòng)物實(shí)驗(yàn)的基本信息:

給藥劑量

mg/kg

動(dòng)物的平均體重

g

每只動(dòng)物的給藥體積

μL

動(dòng)物數(shù)量

由于實(shí)驗(yàn)過(guò)程有損耗,建議您多配一只動(dòng)物的量
計(jì)算結(jié)果
工作液所需濃度 : mg/mL
純度 & 產(chǎn)品資料

純度: 98.87%

參考文獻(xiàn)
Cell Assay
[2]

Hoechst 33258 is prepared as stock solutions in highly pure water. Working solutions in a concentration range of 10-3-10-6 mol/dm3 are prepared prior to testing. Cytotoxic effects of Hoechst 33258 on tested cell lines are determined by the MTT assay. Cells are seeded in 96 micro well flat bottom plates at a concentration of 2×104 cells/mL and left overnight in the CO2 incubator allowing them to attach to the plate surface. Growing medium is replaced with compound supplemented or control medium and incubated for 72 h. Fresh medium with 5 mg/mL of MTT is added onto cells and incubated for 4 h at 37°C. Upon media removal, water insoluble MTT-formazan crystals formed inside the living cells are dissolved in DMSO and the absorbance at 570 nm proportional to the number of living cells is measured on an Elisa Microplate Reader. All experiments are performed at least three times in triplicates.The GI50 value, defined as the compound concentration (μM) leading to cellular growth inhibition by 50%, is calculated and used as a parameter to compare cytotoxicity among the compounds[2].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

參考文獻(xiàn)

完整儲(chǔ)備液配制表

* 請(qǐng)根據(jù)產(chǎn)品在不同溶劑中的溶解度選擇合適的溶劑配制儲(chǔ)備液;一旦配成溶液,請(qǐng)分裝保存,避免反復(fù)凍融造成的產(chǎn)品失效。
儲(chǔ)備液的保存方式和期限:-80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light)。-80°C儲(chǔ)存時(shí),請(qǐng)?jiān)?個(gè)月內(nèi)使用,-20°C儲(chǔ)存時(shí),請(qǐng)?jiān)?個(gè)月內(nèi)使用。

可選溶劑 濃度 溶劑體積 質(zhì)量 1 mg 5 mg 10 mg 25 mg
H2O / DMSO 1 mM 1.8731 mL 9.3654 mL 18.7308 mL 46.8270 mL
5 mM 0.3746 mL 1.8731 mL 3.7462 mL 9.3654 mL
10 mM 0.1873 mL 0.9365 mL 1.8731 mL 4.6827 mL
15 mM 0.1249 mL 0.6244 mL 1.2487 mL 3.1218 mL
20 mM 0.0937 mL 0.4683 mL 0.9365 mL 2.3414 mL
DMSO 25 mM 0.0749 mL 0.3746 mL 0.7492 mL 1.8731 mL
30 mM 0.0624 mL 0.3122 mL 0.6244 mL 1.5609 mL
40 mM 0.0468 mL 0.2341 mL 0.4683 mL 1.1707 mL
50 mM 0.0375 mL 0.1873 mL 0.3746 mL 0.9365 mL
60 mM 0.0312 mL 0.1561 mL 0.3122 mL 0.7805 mL
80 mM 0.0234 mL 0.1171 mL 0.2341 mL 0.5853 mL
100 mM 0.0187 mL 0.0937 mL 0.1873 mL 0.4683 mL

* 備注:如您選擇水作為儲(chǔ)備液,請(qǐng)稀釋至工作液后,再用 0.22 μm 的濾膜過(guò)濾除菌后使用。

Help & FAQs
  • Do most proteins show cross-species activity?

    Species cross-reactivity must be investigated individually for each product. Many human cytokines will produce a nice response in mouse cell lines, and many mouse proteins will show activity on human cells. Other proteins may have a lower specific activity when used in the opposite species.

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產(chǎn)品名稱:
Hoechst 33258 trihydrochloride
目錄號(hào):
HY-15558A
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