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  2. Fluorescent Dye
  3. Hoechst 33258

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

目錄號(hào): HY-15558 純度: 99.11%
COA 產(chǎn)品使用指南 技術(shù)支持

Hoechst 33258 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)。

MCE 的所有產(chǎn)品僅用作科學(xué)研究或藥證申報(bào),我們不為任何個(gè)人用途提供產(chǎn)品和服務(wù)

Hoechst 33258 Chemical Structure

Hoechst 33258 Chemical Structure

CAS No. : 23491-44-3

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  • 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 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
A2780 IC50
12 μM
Compound: HOE-33258
Antitumor activity of compound for 96-h exposure in A2780 human ovarian cell line
Antitumor activity of compound for 96-h exposure in A2780 human ovarian cell line
[PMID: 11170623]
A2780cisR IC50
9.5 μM
Compound: HOE-33258
Antitumor activity of compound for 96-h exposure in A2780cisR (acquired resistance to cisplatin) human ovarian cell line
Antitumor activity of compound for 96-h exposure in A2780cisR (acquired resistance to cisplatin) human ovarian cell line
[PMID: 11170623]
A549 IC50
≥ 31.2 μg/mL
Compound: 2
Cytotoxicity against human A549 cells assessed as reduction in cell survival after 24 hrs by resazurin assay
Cytotoxicity against human A549 cells assessed as reduction in cell survival after 24 hrs by resazurin assay
[PMID: 27301676]
AGS GI50
112 x 10-6 mol/dm3
Compound: Hoechst 33258
Cytotoxicity against human AGS cells after 72 hrs by MTT assay
Cytotoxicity against human AGS cells after 72 hrs by MTT assay
[PMID: 21227551]
BEAS-2B IC50
≥ 31.2 μg/mL
Compound: 2
Cytotoxicity against human BEAS2B cells assessed as reduction in cell survival after 24 hrs by resazurin assay
Cytotoxicity against human BEAS2B cells assessed as reduction in cell survival after 24 hrs by resazurin assay
[PMID: 27301676]
Caco-2 GI50
84 x 10-6 mol/dm3
Compound: Hoechst 33258
Cytotoxicity against human Caco2 cells after 72 hrs by MTT assay
Cytotoxicity against human Caco2 cells after 72 hrs by MTT assay
[PMID: 21227551]
CH1 IC50
26.5 μM
Compound: HOE-33258
Antitumor activity of compound for 96-h exposure in CH1 human ovarian cell line
Antitumor activity of compound for 96-h exposure in CH1 human ovarian cell line
[PMID: 11170623]
HeLa GI50
115.7 x 10-6 mol/dm3
Compound: Hoechst 33258
Cytotoxicity against human HeLa cells after 72 hrs by MTT assay
Cytotoxicity against human HeLa cells after 72 hrs by MTT assay
[PMID: 21227551]
HeLa IC50
51.31 μM
Compound: H. 33258, Hoechst 33258
Cytotoxicity against human HeLa cells after 72 hrs by MTT assay
Cytotoxicity against human HeLa cells after 72 hrs by MTT assay
[PMID: 22959518]
HEp-2 GI50
132.5 x 10-6 mol/dm3
Compound: Hoechst 33258
Cytotoxicity against human Hep2 cells after 72 hrs by MTT assay
Cytotoxicity against human Hep2 cells after 72 hrs by MTT assay
[PMID: 21227551]
HL-60 IC50
32.43 μM
Compound: H. 33258, Hoechst 33258
Cytotoxicity against human HL60 cells after 72 hrs by MTT assay
Cytotoxicity against human HL60 cells after 72 hrs by MTT assay
[PMID: 22959518]
MCF7 GI50
5.7 x 10-6 mol/dm3
Compound: Hoechst 33258
Cytotoxicity against human MCF7 cells after 72 hrs by MTT assay
Cytotoxicity against human MCF7 cells after 72 hrs by MTT assay
[PMID: 21227551]
MDCK GI50
157.5 x 10-6 mol/dm3
Compound: Hoechst 33258
Cytotoxicity against MDCK cells after 72 hrs by MTT assay
Cytotoxicity against MDCK cells after 72 hrs by MTT assay
[PMID: 21227551]
MIA PaCa-2 GI50
191.5 x 10-6 mol/dm3
Compound: Hoechst 33258
Cytotoxicity against human MIAPaCa2 cells after 72 hrs by MTT assay
Cytotoxicity against human MIAPaCa2 cells after 72 hrs by MTT assay
[PMID: 21227551]
NCI-H358 GI50
91.7 x 10-6 mol/dm3
Compound: Hoechst 33258
Cytotoxicity against human NCI-H358 cells after 72 hrs by MTT assay
Cytotoxicity against human NCI-H358 cells after 72 hrs by MTT assay
[PMID: 21227551]
SK-OV-3 IC50
> 100 μM
Compound: HOE-33258
Antitumor activity of compound for 96-h exposure in SKOV-3 human ovarian cell line
Antitumor activity of compound for 96-h exposure in SKOV-3 human ovarian cell line
[PMID: 11170623]
U-937 IC50
15.42 μM
Compound: H. 33258, Hoechst 33258
Cytotoxicity against human U937 cells after 72 hrs by MTT assay
Cytotoxicity against human U937 cells after 72 hrs by MTT assay
[PMID: 22959518]
體外研究
(In Vitro)

Hoechst 工作液的配制
1.1 制備儲(chǔ)存液
用 DMSO 配制 1 mg/mL 的 Hoechst 儲(chǔ)存液。
注:Hoechst 儲(chǔ)存液建議分裝后于 -4℃ 或 -20℃ 避光保存。
1.2 工作液的配制
用預(yù)熱好的無血清細(xì)胞培養(yǎng)基或 PBS 稀釋儲(chǔ)存液,終濃度為 10 μg/mL Hoechst 工作液。
注:請(qǐng)根據(jù)實(shí)際情況調(diào)整 Hoechst 工作液濃度,且現(xiàn)用現(xiàn)配。
2. 細(xì)胞染色(懸浮細(xì)胞)
2.1 離心收集細(xì)胞,加入 PBS 洗滌兩次,每次 5 分鐘。細(xì)胞密度在 1×106/mL
2.2 加入 1 mL Hoechst 工作液,室溫孵育 3-10 分鐘。
2.3 400 g,離心 3-4 分鐘,棄去上清。
2.4 加入 PBS 洗滌細(xì)胞兩次,每次 5 分鐘。
2.5 用 1 mL 無血清培養(yǎng)基或 PBS 重懸細(xì)胞后,使用熒光顯微鏡或流式細(xì)胞儀進(jìn)行觀察。
3. 細(xì)胞染色(貼壁細(xì)胞)
3.1 將貼壁細(xì)胞培養(yǎng)于無菌蓋玻片上。
3.2 從培養(yǎng)基中移出蓋玻片,吸除多余培養(yǎng)基。
3.3 加入 100 μL 染料工作液,輕輕晃動(dòng)使其完全覆蓋細(xì)胞,孵育 3-10 分鐘。
3.4 吸去染料工作液,用培養(yǎng)基洗 2-3 次,每次 5 分鐘,使用熒光顯微鏡或流式細(xì)胞儀進(jìn)行觀察。


注意事項(xiàng)
1. 請(qǐng)根據(jù)實(shí)際情況調(diào)整 Hoechst 工作液濃度。
2. 本產(chǎn)品僅限于專業(yè)人員的科學(xué)研究用,不得用于臨床診斷或治療,不得用于食品或藥品。
3. 為了您的安全和健康,請(qǐng)穿實(shí)驗(yàn)服并戴一次性手套操作。

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

分子量

424.50

Formula

C25H24N6O

CAS 號(hào)
性狀

固體

顏色

Light yellow to yellow

Emission (Em)

451

Excitation (Ex)

356

運(yùn)輸條件

Room temperature in continental US; may vary elsewhere.

儲(chǔ)存方式

-20°C, protect from light

*In solvent : -80°C, 6 months; -20°C, 1 month (protect from light)

溶解性數(shù)據(jù)
細(xì)胞實(shí)驗(yàn): 

DMSO 中的溶解度 : 41.67 mg/mL (98.16 mM; 超聲助溶 (<60°C); 吸濕的 DMSO 對(duì)產(chǎn)品的溶解度有顯著影響,請(qǐng)使用新開封的 DMSO)

配制儲(chǔ)備液
濃度 溶劑體積 質(zhì)量 1 mg 5 mg 10 mg
1 mM 2.3557 mL 11.7786 mL 23.5571 mL
5 mM 0.4711 mL 2.3557 mL 4.7114 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 (protect from 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。

  • 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):

請(qǐng)根據(jù)您的 實(shí)驗(yàn)動(dòng)物和給藥方式 選擇適當(dāng)?shù)娜芙夥桨浮?

以下溶解方案都請(qǐng)先按照 In Vitro 方式配制澄清的儲(chǔ)備液,再依次添加助溶劑:
——為保證實(shí)驗(yàn)結(jié)果的可靠性,澄清的儲(chǔ)備液可以根據(jù)儲(chǔ)存條件,適當(dāng)保存;體內(nèi)實(shí)驗(yàn)的工作液,建議您現(xiàn)用現(xiàn)配,當(dāng)天使用;
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;如在配制過程中出現(xiàn)沉淀、析出現(xiàn)象,可以通過加熱和/或超聲的方式助溶

  • 方案 一

    請(qǐng)依序添加每種溶劑: 10% DMSO    40% PEG300    5% Tween-80    45% Saline

    Solubility: ≥ 2.08 mg/mL (4.90 mM); 澄清溶液

    此方案可獲得 ≥ 2.08 mg/mL(飽和度未知)的澄清溶液。

    1 mL 工作液為例,取 100 μL 20.8 mg/mL 的澄清 DMSO 儲(chǔ)備液加到 400 μL PEG300 中,混合均勻;再向上述體系中加入 50 μL Tween-80,混合均勻;然后再繼續(xù)加入 450 μL 生理鹽水 定容至 1 mL

    生理鹽水的配制:將 0.9 g 氯化鈉,溶解于 ddH?O 并定容至 100 mL,可以得到澄清透明的生理鹽水溶液。
  • 方案 二

    請(qǐng)依序添加每種溶劑: 10% DMSO    90% (20% SBE-β-CD in Saline)

    Solubility: ≥ 2.08 mg/mL (4.90 mM); 澄清溶液

    此方案可獲得 ≥ 2.08 mg/mL(飽和度未知)的澄清溶液。

    1 mL 工作液為例,取 100 μL 20.8 mg/mL 的澄清 DMSO 儲(chǔ)備液加到 900 μL 20% 的 SBE-β-CD 生理鹽水水溶液 中,混合均勻。

    2 g SBE-β-CD(磺丁基醚 β-環(huán)糊精)粉末定容于 10 mL 的生理鹽水中,完全溶解至澄清透明。
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)過程有損耗,建議您多配一只動(dòng)物的量
請(qǐng)輸入您的動(dòng)物體內(nèi)配方組成:
%
DMSO +
+
%
Tween-80 +
%
Saline
如果您的動(dòng)物是免疫缺陷鼠或者體弱鼠,建議 DMSO 中的在最后工作液體系中的占比盡量不超過 2%。
方案所需 助溶劑 包括:DMSO, ,均可在 MCE 網(wǎng)站選購。 ,Tween 80,均可在 MCE 網(wǎng)站選購。
計(jì)算結(jié)果
工作液所需濃度 : mg/mL
儲(chǔ)備液配制方法 : mg 藥物溶于 μL  DMSO(母液濃度為 mg/mL)。

*In solvent : -80°C, 6 months; -20°C, 1 month (protect from light)

您所需的儲(chǔ)備液濃度超過該產(chǎn)品的實(shí)測(cè)溶解度,以下方案僅供參考,如有需要,請(qǐng)與 MCE 中國技術(shù)支持聯(lián)系。
動(dòng)物實(shí)驗(yàn)體內(nèi)工作液的配制方法 : 取 μL DMSO 儲(chǔ)備液,加入 μL  μL ,混合均勻至澄清,再加 μL Tween 80,混合均勻至澄清,再加 μL 生理鹽水。
連續(xù)給藥周期超過半月以上,請(qǐng)謹(jǐn)慎選擇該方案。
請(qǐng)確保第一步儲(chǔ)備液溶解至澄清狀態(tài),從左到右依次添加助溶劑。您可采用超聲加熱 (超聲清洗儀,建議頻次 20-40 kHz),渦旋吹打等方式輔助溶解。
純度 & 產(chǎn)品資料

純度: 99.93%

染色示例
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 (protect from 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
DMSO 1 mM 2.3557 mL 11.7786 mL 23.5571 mL 58.8928 mL
5 mM 0.4711 mL 2.3557 mL 4.7114 mL 11.7786 mL
10 mM 0.2356 mL 1.1779 mL 2.3557 mL 5.8893 mL
15 mM 0.1570 mL 0.7852 mL 1.5705 mL 3.9262 mL
20 mM 0.1178 mL 0.5889 mL 1.1779 mL 2.9446 mL
25 mM 0.0942 mL 0.4711 mL 0.9423 mL 2.3557 mL
30 mM 0.0785 mL 0.3926 mL 0.7852 mL 1.9631 mL
40 mM 0.0589 mL 0.2945 mL 0.5889 mL 1.4723 mL
50 mM 0.0471 mL 0.2356 mL 0.4711 mL 1.1779 mL
60 mM 0.0393 mL 0.1963 mL 0.3926 mL 0.9815 mL
80 mM 0.0294 mL 0.1472 mL 0.2945 mL 0.7362 mL
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
目錄號(hào):
HY-15558
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