Recombinant Human Granulocyte-Colony Stimulating Factor,CHO
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英文名稱:Recombinant Human Granulocyte-Colony Stimulating Factor,CHO
中文簡稱:【G CSF CHO】重組人粒細(xì)胞集落刺激因子,CHO表達(dá)
Recombinant Human Granulocyte-Colony Stimulating Factor,CHO 【G CSF CHO】重組人粒細(xì)胞集落刺激因子,CHO表達(dá) 細(xì)胞因子產(chǎn)品描述:
粒細(xì)胞集落刺激因子(granulocyte colony stimulating factor,G-CSF)基因全長2.5kb,包括5個(gè)外顯子和4個(gè)內(nèi)含子,人G-CSF基因位于17號(hào)染色體,與小鼠G-CSF基因約有73%同源性,與IL-6無論在基因水平以及氨基酸水平上都有很高同源性,包括外顯子、內(nèi)含子組成,Cys數(shù)目,兩對(duì)二硫鍵位置以及分子的三級(jí)結(jié)構(gòu)等。G-CSF主要作用于中性粒細(xì)胞系造血細(xì)胞的增殖、 分化和活化。在體外G-CSF刺激骨髓造血祖細(xì)胞中中性粒細(xì)胞集落的形成,延長成熟中性粒細(xì)胞的存活時(shí)間,活化中性粒細(xì)胞如促進(jìn)ADCC,超氧陰離子的產(chǎn)生和堿性磷酸酶的合成。G-CSF還具有對(duì)人粒細(xì)胞、單核細(xì)胞、成纖維細(xì)胞、平滑肌細(xì)胞以及成肌纖維細(xì)胞的趨化作用。腫瘤患者注射G-CSF后可提高血循環(huán)中中性粒細(xì)胞的水平,這種作用可能與縮短某些骨髓細(xì)胞進(jìn)入S期的時(shí)間以及增加生成粒細(xì)胞的祖細(xì)胞數(shù)量有關(guān)。
人粒細(xì)胞集落刺激因子,CHO表達(dá)重組蛋白是在CHO中國倉鼠卵巢細(xì)胞中重組表達(dá)的,是一條單鏈、糖基化的多肽鏈,包含有174個(gè)氨基酸,分子量為18kDa。該人粒細(xì)胞集落刺激因子,CHO表達(dá)重組蛋白最終由標(biāo)準(zhǔn)的色譜技術(shù)純化出。
來源宿主---CHO中國倉鼠卵巢細(xì)胞中重組表達(dá)
Recombinant Human Granulocyte-Colony Stimulating Factor,CHO 【G CSF CHO】重組人粒細(xì)胞集落刺激因子,CHO表達(dá) 細(xì)胞因子使用須知:
溶解:提供的人粒細(xì)胞集落刺激因子,CHO表達(dá)(G CSF CHO)產(chǎn)品為過濾滅菌的白色凍干粉末;該凍干粉末是從含1mg/ml人粒細(xì)胞集落刺激因子,CHO表達(dá)(G CSF CHO)的10mM鹽酸(pH 6.5),0.4mg Tween 20,100mg甘露醇,160 mg左型精氨酸,40 mg苯基丙氨酸和4mg甲硫氨酸中凍干產(chǎn)生的。我們建議加無菌的18MΩ-cm水以配制工作儲(chǔ)存液到100ug/ml,后繼實(shí)驗(yàn)過程中可根據(jù)實(shí)驗(yàn)要求進(jìn)一步稀釋。
保存:人粒細(xì)胞集落刺激因子,CHO表達(dá)(G CSF CHO)凍干粉末在室溫可保存3周,或置于-18℃凍存用于長期保存。重新溶解后可以在4℃冷藏保存至少2-7天以上;如果想延長保存時(shí)間,建議分裝(加載體蛋白如0.1% HSA或 BSA)后置于-18℃凍存。請(qǐng)避免反復(fù)凍融。
Recombinant Human Granulocyte-Colony Stimulating Factor,CHO 【G CSF CHO】重組人粒細(xì)胞集落刺激因子,CHO表達(dá) 細(xì)胞因子英文描述:
Synonyms
SF-3, MGI-1G, GM-CSF beta, Pluripoietin, Filgrastim, Lenograstim, G-CSF, MGC45931, GCSF.
Introduction
Granulocyte Colony Stimulating Factor is a growth factor and/or cytokine produced by the endothelium, macrophages and a number of other immune cells. GCSF stimulates the bone marrow to produce granulocytes and also to stimulate the survival, proliferation, differentiation and function of neutrophil granulocyte progenator cells and mature neutrophils.
Description
Granulocyte Colony Stimulating Factor Human Recombinant produced in CHO cells is a single, glycosylated, polypeptide chain containing 174 amino acids and having a molecular mass of approximately 18 kDa.
G-CSF is purified by proprietary chromatographic techniques.
Source (來源)
Chinese Hamster Ovary Cells (CHO).
Physical Appearance (物理特征)
Sterile Filtered White lyophilized (freeze-dried) powder.
Formulation
G-CSF was lyophilized from a concentrated (1mg/ml) Phosphate- Buffered Saline, pH 7.4.
Solubility (溶解性)
It is recommended to reconstitute the lyophilized Granulocyte Colony Stimulating Factor in sterile 18MΩ-cm H2O not less than 100µg/ml, which can then be further diluted to other aqueous solutions.
Stability (穩(wěn)定性)
Lyophilized Granulocyte Colony Stimulating Factor although stable at room temperature for 3 weeks, should be stored desiccated below -18°C. Upon reconstitution G-CSF should be stored at 4°C between 2-7 days and for future use below -18°C.
For long term storage it is recommended to add a carrier protein (0.1% HSA or BSA).
Please prevent freeze-thaw cycles.
Amino acid sequence (氨基酸序列)
TPLGPASSLP QSFLLKCLEQ VRKIQGDGAA LQEKLCATYK LCHPEELVLL GHSLGIPWAP LSSCPSQALQ LAGCLSQLHS GLFLYQGLLQ ALEGISPELG PTLDTLQLDV ADFATTIWQQ MEELGMAPAL QPTQGAMPAF ASAFQRRAGG VLVASHLQSF LEVSYRVLRH LAQP.
Purity (純度)
Greater than 97.0% as determined by:
a) Analysis by RP-HPLC.
b) Analysis by SDS-PAGE.
Biological Activity (生物活性)
The ED50, calculated by the dose-dependant proliferation of murine NFS-60 indicator cells (measured by 3H-thymidine uptake) is < 0.07 ng/ml, corresponding to a Specific Activity of 1.27 x 108 IU/mg.
Protein content (蛋白質(zhì)含量)
GCSF quantitation was carried out by two independent methods:
1) UV spectroscopy at 280 nm using the absorbency value of 0.815 as the extinction coefficient for a 0.1% (1mg/ml) solution. This value is calculated by the PC GENE computer analysis program of protein sequences (IntelliGenetics).
2) Analysis by RP-HPLC, using a calibrated solution of Granulocyte Colony Stimulating Factor as a Reference Standard.
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