Recombinant Human Interferon-gamma
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英文名稱:Recombinant Human Interferon-gamma
中文簡(jiǎn)稱:【IFN g】重組人干擾素-γ
Recombinant Human Interferon-gamma 【IFN g】重組人干擾素-γ 細(xì)胞因子描述:
干擾素(interferon,IFN)是細(xì)胞對(duì)相關(guān)刺激反應(yīng)時(shí)所產(chǎn)生的細(xì)胞信號(hào)蛋白質(zhì),是細(xì)胞因子超家族中一個(gè)糖蛋白類同源細(xì)胞因子家族,稱為IFN家族。已鑒定出該家族中的3個(gè)主要成員:IFN-α、IFN-β和IFN-γ。其中干擾素-γ(Interferon-gamma,IFN-γ)是一種具有抗病毒、抗腫瘤和免疫調(diào)節(jié)作用的細(xì)胞因子,主要由活化的T細(xì)胞和NK細(xì)胞產(chǎn)生,在機(jī)體免疫系統(tǒng)中發(fā)揮重要作用。IFN-γ是一類分泌型糖蛋白,由143個(gè)氨基酸組成,相對(duì)分子量是16924。而重組人干擾素-γ相對(duì)分子量在14000-16000之間,這是由于重組IFN-γ在C末端有氨基酸丟失現(xiàn)象。
人干擾素-γ重組蛋白是在大腸桿菌中重組表達(dá)的,是一條單鏈、非糖基化的多肽鏈,包含有144個(gè)氨基酸,分子量為16879Da。該人干擾素-γ重組蛋白最終由標(biāo)準(zhǔn)的色譜技術(shù)純化出。
大腸桿菌中重組表達(dá)
Recombinant Human Interferon-gamma 【IFN g】重組人干擾素-γ 細(xì)胞因子實(shí)驗(yàn)須知:
溶解:提供的人干擾素-γ(IFN g)產(chǎn)品為過(guò)濾滅菌的白色凍干粉末;該凍干粉末是從含1mg/ml人干擾素-γ(IFN g)的10mM磷酸鈉緩沖液(pH 7.4±0.1)中經(jīng)進(jìn)一步透析并凍干產(chǎn)生的。我們建議加無(wú)菌的18MΩ-cm水以配制工作儲(chǔ)存液到100ug/ml,后繼實(shí)驗(yàn)過(guò)程中可根據(jù)實(shí)驗(yàn)要求進(jìn)一步稀釋。
保存:人干擾素-γ(IFN g)凍干粉末在室溫可保存3周,或置于-18℃凍存用于長(zhǎng)期保存。重新溶解后可以在4℃冷藏保存至少2-7天;如果想延長(zhǎng)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。
蛋白含量:蛋白質(zhì)定量是由兩個(gè)獨(dú)立的方法
1。紫外光譜在280海里使用的吸墨性價(jià)值0.640的消光系數(shù)0.1%(1毫克/毫升)的解決方案。這個(gè)值是由電腦計(jì)算基因蛋白質(zhì)序列的計(jì)算機(jī)分析程序(IntelliGenetics)。
2。產(chǎn)物的分析,使用校準(zhǔn)IFN-g作為參考標(biāo)準(zhǔn)的解決方案。
Recombinant Human Interferon-gamma 【IFN g】重組人干擾素-γ 細(xì)胞因子英文描述:
Synonyms
Immune Interferon, type II interferon, T cell interferon, MAF, IFNG, IFG, IFI, IFN-gamma.
Introduction
IFN-gamma produced by lymphocytes activated by specific antigens or mitogens.
IFN-gamma, in addition to having antiviral activity, has important immunoregulatory functions, it is a potent activator of macrophages, and has antiproliferative effects on transformed cells and it can potentiate the antiviral and antitumor effects of the type I interferons.
Description
Interferon-gamma Human Recombinant produced in E.Coli is a single, non-glycosylated, polypeptide chain containing 144 amino acids and having a molecular mass of 17kDa.
The IFN-gamma is purified by proprietary chromatographic techniques.
Source
Escherichia Coli.
Physical Appearance
Sterile Filtered White lyophilized (freeze-dried) powder.
Formulation
The protein was lyophilized from a 0.2µm filtered concentrated solution in PBS pH 7.4.
Solubility
It is recommended to reconstitute the lyophilized Interferon-gamma in sterile 18MΩ-cm H2O not less than 100µg/ml, which can then be further diluted to other aqueous solutions.
Stability
Lyophilized Interferon gamma although stable at room temperature for 3 weeks, should be stored desiccated below -18°C. Upon reconstitution IFN-gamma 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.
Purity
Greater than 98.0% as determined by:
(a) Analysis by RP-HPLC.
(b) Analysis by SDS-PAGE.
Amino acid sequence
MQDPYVKEAE NLKKYFNAGH SDVADNGTLF LGILKNWKEE SDRKIMQSQI VSFYFKLFKN FKDDQSIQKS VETIKEDMNV KFFNSNKKKR DDFEKLTNYS VTDLNVQRKA IHELIQVMAE LSPAAKTGKR KRSQMLFQGR RASQ.
Biological Activity
The specific activity as determined in a viral resistance assay is < 0.05 ng/ml, corresponding to a specific activity of 2.0 x 10,000,000 IU/mg.
Protein content
Protein quantitation was carried out by two independent methods:
1. UV spectroscopy at 280 nm using the absorbency value of 0.640 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 IFN-g as a Reference Standard.
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