Zeocin™
Catalog no.  R250
Quantity 0.125 g (8 × 1.25 ml)
Store at -20°C

貨號(hào)：R250-01（原裝貨號(hào)）/Z8020（分裝貨號(hào)）
規(guī)格：125mg/1.25ml/1G
式量： 1137.41 克/摩爾
濃度/純度：100 mg/mL
分子式/化學(xué)式:C55H83N19O21S2Cu
分子量：1137.41
外觀（性狀）：淡綠色液體
儲(chǔ)存條件： -20℃避光
From Invitrogen R250-01

Description
Zeocin™ is a formulation of phleomycin D1, a basic, water-soluble, copperchelated glycopeptide isolated fromStreptomyces verticillus and shows strong toxicity against bacteria, fungi (including yeast), plants, and mammalian cell
lines. The blue color of the solution is due to the presence of copper and the copper-chelated form of Zeocin™ is inactive.
When the antibiotic enters the cell, the copper cation is reduced from Cu2+ to Cu1+ and removed by sulfhydrylcompounds in the cell. Upon copper removal, Zeocin™ is activated, and binds and cleaves DNA, causing cell death. A
Zeocin™ resistance protein of 13,665 Da, has been isolated and characterized. The protein is the product of the Sh ble
gene (Streptoalloteichus hindustanus bleomycin gene), binds stoichiometrically to Zeocin™ and inhibits its DNA strand
cleavage activity. Expression of this protein in eukaryotic and prokaryotic hosts confers resistance to Zeocin™.
Note: Basic information on using Zeocin™ is described in this insert. Fordetails including Zeocin™ structure and
photographs of Zeocin™ treated cells, download the Zeocin™ manual from www.invitrogen。。ｃｏｍ.
Specifications
Contents: 100 mg/ml solution in deionized, autoclaved water.
Shipping/Storage: Shipped on blue ice. Store at -20°C.
E. coli Selection: 25-50 µg/ml in low salt LB medium (NaCl concentration should not exceed 5 g/liter.)
Yeast Selection: 50-300 µg/ml in YPD or minimal medium
Mammalian Cells 50-1000 µg/ml in suitable medium (varies with cell Selection: line).
Handling Zeocin™
• Always wear gloves, a laboratory coat, and safety glasses when handling Zeocin™ containing solutions.
• Zeocin™ is light sensitive. Store the antibiotic and plates or medium containing the antibiotic in the dark.
• Reduce the salt in bacterial medium and adjust the pH to 7.5 to keep Zeocin™ active as high ionic strength and acidity
or basicity inhibit Zeocin™ activity.
• Store Zeocin™ at -20℃ and thaw on ice before use.
Zeocin™ Selection in E. coli
Host: Must not contain the Tn5 transposon (i.e. *0, DH5, DH10).
Medium: Use Low Salt LB Medium (10 g Tryptone, 5 g NaCl, and 5 g Yeast Extract) at pH 7.5 to prevent inactivation of
Zeocin™.
Selection: Use 25-50 µg/ml of Zeocin™ for selection in E. coli.
Zeocin™ Selection in Yeast
Yeast: Saccharomyces cerevisiae, Pichia pastoris
Medium: YPD with 1 M sorbitol (electroporated cells); YPD or minimal plates (chemically transformed cells). Test the
medium adjusted to pH values ranging from 6.5-8.0 and select the pH that allows you to use lowest
Zeocin™ concentration.
Transformation Method: Use electroporation, lithium cation protocols, or EasyComp™ Kits. Do not use spheroplasting
for yeast transformation with Zeocin™ containing plasmids as it results in complete cell death.
Selection: Use 50-300 µg/ml of Zeocin™, depending on the yeast strain, and media pH and ionic strength. Perform a kill
curve to determine the lowest Zeocin concentration required to kill the untransformed host strain.
Note: Allow the cells to recover for 1 hour in YPD medium after transformation. To obtain efficient Zeocin™ selection,
plate at low cell densities (use 10, 25, 50, 100, and 200 µl of transformation reaction).
Zeocin™ Selection in Mammalian Cells
Use 50-1000 µg/ml of Zeocin™ to select stable cell lines (the average is about 250-400 µg/ml). Depending on the cell line,
ittakes 2-6 weeks to generate foci with Zeocin™. Determine the minimum concentration required to kill your
untransfected host cell line prior to generating stable cell lines (see below).
Determining Zeocin™ Sensitivity
1. Plate or split a confluent plate to obtain cells at ~25% confluency. Prepare a set of 8 plates. Grow cells for 24 hours.
Remove the medium.
2. Add medium with varying Zeocin™ concentrations (0, 50, 100, 200, 400, 600, 800, and 1000 µg/ml) to each plate.
3. Replenish selective medium every 3-4 days and observe the percentage of surviving cells. Select the concentration
that kills the majority of cells within 1-2 weeks.
Selecting Stable Integrants
1. Transfect your cell line and plate onto 100 mm culture plates. Include a sample of untransfected cells as a negative
control.
2. After transfection, wash the cells once with 1X PBS and add fresh medium to the cells.
3. Forty-eight to 72 hours after transfection, split the cells using various dilutions into fresh medium containing Zeocin™
at the pre-determined concentration required for your cell line. To have a better chance at identifying and selecting foci,
we recommend using different cell dilutions.
4. Feed the cells with selective medium every 3-4 days until cell foci are identified.
5. Pick and transfer colonies to 96- or 48-well plates. Grow cells to near confluence before expanding to larger wells or
plates.
Zeocin™ Selection in Mammalian Cells, Continued
Selection Tip
If your cells are more resistant to Zeocin™, split cells into medium containing Zeocin™ and incubate the cells at 37℃ for
2-3 hours to let cells attach. Place the cells at 4℃ for 2 hours. Remember to buffer the medium with HEPES. Return the
cells to 37°C.
Incubating the cells at 4°C stops the cell division process for a short time, allowing Zeocin™ to act, resulting in cell death.
Maintaining Stable Cell Lines
• Maintain cells in the same Zeocin™ concentration used for selection
• Reduce the Zeocin™ concentration by half or to a concentration that just prevents growth of sensitive cells but does not
kill them (refer to the kill curve experiment)
Product Qualification
Zeocin™ is lot qualified by demonstrating that LB media containing 25 µg/ml Zeocin™ prevents growth of the E. coli
strain, *0.

說(shuō)明: Zeocin是一種屬于爭(zhēng)光霉素家族的糖蛋白抗生素， 在體內(nèi)能作用于大多數(shù)細(xì)菌(包括E. coli)、真菌(如：酵母菌)、植物細(xì)、動(dòng)物細(xì)胞。

產(chǎn)品描述：

    Zeocin試劑是博萊霉素抗生素家族的一個(gè)成員。 其抗性由編碼一種 13,665 道爾頓蛋白質(zhì)的 Sh ble 基因賦予。 在表達(dá)這種蛋白質(zhì)的細(xì)胞中，Zeocin無(wú)法結(jié)合和切割細(xì)胞 DNA。 Zeocin對(duì)大多數(shù)好氧細(xì)胞有效，可用于選擇哺乳動(dòng)物和昆蟲細(xì)胞系、酵母以及細(xì)菌。 選擇細(xì)胞時(shí)，Zeocin的用量為 50-2000 ug/ml（通常 300 ug/ml），具體取決于細(xì)胞類型。

    Zeocin 可選擇 Sh ble 基因表達(dá)的細(xì)胞。與現(xiàn)在使用的其他動(dòng)物細(xì)胞標(biāo)記物無(wú)交叉抗性。因此這種抗生素可用來(lái)分離對(duì)其他篩選劑 ( 如：慶大霉素，潮霉素 ) 有抗性的克隆。 Zeocin 是一種屬于爭(zhēng)光霉素家族的糖蛋白抗生素，在體內(nèi)能作用于大多數(shù)細(xì)菌 ( 包括 E 。 coli) 、真菌 ( 如：酵母菌 ) 、植物細(xì)胞、動(dòng)物細(xì)胞、細(xì)胞培養(yǎng)級(jí)。

博萊霉素/Zeocin
     抗生素是微生物的代謝產(chǎn)物，是由真菌、細(xì)菌或其他生物在繁殖過(guò)程中所產(chǎn)生的一類具有殺滅或抑制微生物生長(zhǎng)的物質(zhì)，也可用人工合成的方法制造，用很小的劑量就能抑制或殺滅病原微生物。自1940年青霉素應(yīng)用于臨床以來(lái)，抗生素的種類已達(dá)幾千種，在臨床上常用的亦有幾百種，主要是從微生物的培養(yǎng)液中提取的或者用合成、半合成方法制造，分類有以下幾種：
（一）β-內(nèi)酰胺類
      青霉素類和頭孢菌素類的分子結(jié)構(gòu)中含有β-內(nèi)酰胺環(huán)。近年來(lái)又有較大發(fā)展，如硫酶素類（thienamycins）、單內(nèi)酰環(huán)類(monobactams)，β-內(nèi)酰酶抑制劑(β-lactamadeinhibitors)、甲氧青霉素(methoxypeniciuins)等。
（二）氨基糖甙類
包括鏈霉素、慶大霉素、卡那霉素、妥布霉素、丁胺卡那霉素、新霉素、核糖霉素、小諾霉素、阿斯霉素等。
（三）四環(huán)素類
包括四環(huán)素、土霉素、金霉素及多西環(huán)素素等。
（四）氯霉素類
包括氯霉素、甲砜霉素等。
（五）大環(huán)內(nèi)脂類
臨床常用的有紅霉素、白霉素、無(wú)味紅霉素、琥乙紅霉素、阿奇霉素、乙酰螺旋霉素、麥迪霉素、交沙霉素等。
（六）作用于G+細(xì)菌的其它抗生素
如林可霉素、氯林可霉素、萬(wàn)古霉素、桿菌肽等。
（七）作用于G菌的其它抗生素
如多粘菌素、磷霉素、卷霉素、環(huán)絲氨酸、利福平等。
（八）抗真菌抗生素
如灰黃霉素。
（九）抗腫瘤抗生素
如放線菌素D、博萊霉素、阿霉素等。
（十）具有免疫抑制作用的抗生素 如環(huán)孢素。

Zeocin™訂購(gòu)說(shuō)明：
1、絕大部分產(chǎn)品備有現(xiàn)貨，一般情況下都能訂貨當(dāng)日發(fā)貨。 
2、部分非常用產(chǎn)品，需提前1－2日預(yù)訂，海外期貨則需要提前3-6周預(yù)訂。 
3、部分產(chǎn)品價(jià)格會(huì)因貨期、批次等因素發(fā)生變化，若有變動(dòng)以訂貨當(dāng)日新價(jià)格為準(zhǔn)。 
4、每日訂單截止時(shí)間為16點(diǎn)整，部分城市可到17點(diǎn)整，因超過(guò)截止時(shí)間造成當(dāng)日不能發(fā)貨的將于次日安排發(fā)貨。
5、請(qǐng)辦理完貨款后，將收據(jù)、底單等連同收貨人的地址、姓名、等以傳真、、短信、等形式通知我們。

Zeocin™運(yùn)輸說(shuō)明：
1、極低溫產(chǎn)品：極低溫產(chǎn)品運(yùn)輸過(guò)程中加裝干冰運(yùn)輸。用干冰把產(chǎn)品包裹起來(lái)，再用泡沫盒密封，膠帶層層粘住泡沫盒，放入索萊寶的箱子，然后交付給合作快遞，安全快速高效放心的送客戶的手中，保證產(chǎn)品的性質(zhì)穩(wěn)定如一，為您的實(shí)驗(yàn)保駕護(hù)航。
2、低溫產(chǎn)品：低溫產(chǎn)品運(yùn)輸過(guò)程中加裝索萊寶冰袋運(yùn)輸。事先用冰袋把產(chǎn)品包裹起來(lái)，再使用泡沫盒密封，用膠帶嚴(yán)實(shí)密封泡沫盒，再放入索萊寶的箱子（保溫效果少可以持續(xù)一周），然后交付給合作快遞，安全快速高效放心的送客戶的手中，保證產(chǎn)品的性質(zhì)穩(wěn)定如一，為您的實(shí)驗(yàn)保駕護(hù)航。
3、常溫產(chǎn)品：常溫產(chǎn)品運(yùn)輸過(guò)程中無(wú)需加冰或者特殊包裝。產(chǎn)品由公司庫(kù)房人員快速配貨，準(zhǔn)確快速高效的快遞保證產(chǎn)品快速送達(dá)您的手中。