成人欧美一区二区三区黑人免费_亚洲永久无码7777KKK_国产欧美日韩综合精品一区二区_欧美在线一区二区三区

English | 中文版 | 手機版 企業(yè)登錄 | 個人登錄 | 郵件訂閱
當前位置 > 首頁 > 技術文章 > 新的分子間相互作用分析儀微量熱泳動儀---MST 用戶評價舉例

新的分子間相互作用分析儀微量熱泳動儀---MST 用戶評價舉例

瀏覽次數:30686 發(fā)布日期:2013-4-8  來源:本站 僅供參考,謝絕轉載,否則責任自負
新的分子間相互作用分析儀
微量熱泳動儀---MST
用戶評價舉例

德國Nano Temper公司
南開大學元素有機化學國家重點實驗室
南開大學化學學院副院長
南開大學化學生物學系主任
席真
研究方向:蛋白質-蛋白質
http://skleoc.nankai.edu.cn/professors/xiz/index.html
 
Our recent research is focused on the molecular basis of herbicide resistance and exploiting new target sites of herbicide, such as PPI of AHAS. Via MST, we measured the Protein-Protein interaction, that have been difficult to detect and quantify with ITC because of instability of the protein and very little heat changes of our samples. The MST technology can be used as a complementary technology to the traditional methods in these fields.
We can strongly recommend the MST technology for its low material consumption, short measurement times and broad application range in the field of molecule interaction studies.
我們目前致力于除草劑抗性的分子機制以及尋找新的除草劑靶點(比如AHAS亞基間相互作用的界面)等方面的研究。通過MST,我們測量了蛋白質-蛋白質相互作用,而我們的樣品使用ITC方法測定沒有得到結果,因為我們的樣品穩(wěn)定性較差并且相互作用熱量變化極小。所以在這些研究領域,MST技術能夠很好地與傳統(tǒng)的技術互補。
我們非常推薦MST技術的一些特點,如低樣品使用量、很短的測量時間以及在分子間相互作用方面很廣泛的應用。
生物物理學研究所
生物物理學研究所分子生物學研究中心主任
中國科學院院士
王大成
研究方向:蛋白質-DNA
http://sourcedb.cas.cn/sourcedb_ibp_cas/
 
Our resent research is focused on the critical pathogenic protein & complex from pathogenic microbes and The Protein structure-functional basis for some endogenous disease. Via MST, we measured the Protein-DNA interaction, that have been difficult to detect and quantify with other standard biomolecule interaction technologies (like SPR and ITC) because of the conformation/immobilization-sensitive and very little heat changes of our samples. MST technology can be used as a complementary technology to the traditional methods in these fields.
We can strongly recommend MST technology for its low material consumption, short measurement times and broad application range for molecule-interaction studies.
我們目前致力于重要病原菌致病關鍵蛋白質的結構-功能和分子機制和一些內源性疾病的蛋白質結構-功能基礎等方面的研究。通過MST,我們測量了蛋白質-DNA相互作用,而我們的樣品很難通過其他傳統(tǒng)方法(例如SPR和ITC)來測量,因為我們的樣品構象對固定很敏感并且熱量變化極小。所以在這些研究領域,MST技術能夠很好地與傳統(tǒng)的技術互補。
我們非常推薦MST技術的一些特點,如低樣品使用量、很短的測量時間以及在分子間相互作用方面很廣泛的應用。
山東大學生命科學學院
山東大學微生物技術國際重點實驗室博士生導師
985平臺學術帶頭人
谷立川
研究方向:蛋白質-小分子抑制劑,蛋白質結構與功能
http://www.lifegrad.sdu.edu.cn/a/boshishengdaoshi/2010/0104/151.html
 
Part of our research focus on the protein structural-function basis for iron metabolism of pathogenic microorganisms. We measured the dissociated constant for periplasmic binding protein and siderophore through MST technology. Because siderophore is extremely insoluble, it's difficult for us to complete that task via other methods such as ITC and SPR. We strongly recommend MST technology for its low material consumption, short measurement time and very wide application range.
(編者譯)
我們目前一部分課題是關于致病微生物鐵代謝相關蛋白的結構與功能。通過MST技術,我們測量了周質空間轉鐵蛋白與鐵載體的解離常數,我們的樣品很難通過其他傳統(tǒng)方法(例如SPR和ITC)來測量因為小分子的溶解度極低,MST技術的我們得到了理想的結果。我們非常推薦MST技術的一些特點,如極低的樣品使用量,較快的測量時間,以及十分廣泛的應用
法國賽諾菲公司研發(fā)部
www.sanofi.com
用戶:Dr. Alexey Rak
研究方向:藥物開發(fā)
 
“We routinely assess interaction affinity for both small molecule and biologics projects, with NanoTemper Technologies’ Microscale Thermophoresis being the most recent addition to the pool of instruments we use to carry out these measurements. It has proved a valuable tool for characterising small molecule-protein and protein-protein interactions, as well as for the study of protein aggregation concentration determination. There is very good agreement with other technologies such as Surface Plasmon Resonance (SPR) and Isothermal Titration Calorimetry (ITC), and we are particularly appreciative of this new technology because of the extremely low protein consumption and relatively short time required for the assay setup. NanoTemper customer support has been a key factor in enabling us to familiarise ourselves with the new technology. We would like to deploy increasing numbers of applications based on MST technologies and continue to interact with NanoTemper Technologies Company, a dynamic, scientifically driven company.”
(編者譯)
我們經常需要測量小分子和生物制劑的相互作用親和力,我們最近使用的最新一個設備就是NanoTemper的微量熱泳動技術。結果證明了MST是一個有價值的工具,用于測量小分子-蛋白質、蛋白質之間的相互作用,以及判斷蛋白質形成聚集體的濃度。MST技術與傳統(tǒng)的其他技術如表面等離子體共振(SPR)和等溫滴定量熱法(ITC)的結果完全吻合,我們特別喜歡這個新技術的一些特點:極低的蛋白樣品用量和很短的實驗設置時間。NanoTemper的技術支持非常及時有效,使我們能夠很快熟悉這個新技術。我們很愿意使用越來越多MST儀器并繼續(xù)與NanoTemper技術公司相互合作,因為這個一個積極向上的、技術先進的公司。
德國馬爾堡菲利普大學,藥物化學研究所
網站:http://www.agklebe.de/
用戶:Prof. Gerhard Klebe
研究方向:結構性藥物設計
 
 
The main focus of our work lies on structure based-drug design. In this context, we study the interaction of the bacterial tRNA modifying enzyme tRNA-guanine transglycosylase (Tgt) with small molecule inhibitors and with tRNA. Although we were unable to measure any interaction of this enzyme with its small molecule substrates guanine and 7-deaza-7-aminomethyl guanine via MST, a strong influence on the thermophoretic behaviour of Tgt was noticed upon binding of the Macromolecular tRNA substrate allowing determination of the respective Kd value.In addition, MST yielded Kd values for lin-benzoguanine-based Tgt inhibitors (300-500 Da), which were excellently consistent with Ki values previously determined in enzyme kinetic studies. Accordingly, this fast and easy to use method provides a highly welcome alternative to the radioactive enzyme assay we used so far to figure out binding affinities of Tgt inhibitors. Furthermore, we investigate the interaction of a chaperone of the Shigella type III secretion system with its protein interaction partners or rather with synthetic peptides thereof to identify the respective recognition motifs. Via MST, we determined with low amounts of protein material Kd values which were in nearly perfect agreement with those measured via isothermal titration calorimetry.
(編者譯)
我們的主要工作是根據結構設計藥物。比如,細菌tRNA修飾酶-tRNA鳥嘌呤轉糖苷酶(Tgt)與小分子抑制劑以及tRNA的相互作用。雖然我們無法直接衡量這個酶的與小分子鳥嘌呤以及7-deaza-7-aminomethyl的相互作用,但是由于Tgt結合大分子tRNA是有很明顯的熱泳動效應變化,我們可以通過MST來測量各自的Kd值。此外,MST測量出來lin-benzoguanine-based MST Tgt抑制劑(300 - 500 Da) 的Kd值極好地吻合先前確定通過酶動力學研究得到的Ki值。因此,這個快速和易于使用的方法提供了一個非常好的方法用于替代放射性酶活性測定,而后者是我們之前一直用來測量Tgt抑制劑親和力的方法。我們使用到目前為止找出綁定的親和力Tgt抑制劑。此外,我們還研究了一個志賀氏桿菌III型分泌系統(tǒng)分子伴侶蛋白與其結合物(合成的多肽)的相互作用。通過MST,我們能通過極低量的蛋白質來測定Kd值,而且其結果幾乎符合傳統(tǒng)的等溫滴定量熱法ITC等技術所得到的數據。
法國國家科研中心
http://seg.ihes.fr/
用戶:Dr. Arndt Benecke
研究方向:小非編碼RNA
 
We use systems biology to study transcription regulatory phenomena on a genome-wide scale in the context of host-pathogen interactions and cancer.
Microscale Thermophoresis (MST) thereby has been extremely useful to rapidly quantify relevant protein-nucleic acid and protein-protein interactions directly in cellular lysates without going through the hassles of purification.
(編者譯)
我們用系統(tǒng)生物學方法在全基因組范圍研究轉錄調控現(xiàn)象,研究對象是癌癥和宿主—病原體。微量熱泳動儀(MST)非常有價值的方法,因為其快速量化定量蛋白質-核酸和蛋白質之間的相互作用,而且不需要復雜繁瑣的純化即在細胞裂解液之中完成測量
美國安德森癌癥中心
http://faculty.mdanderson.org/John_Ladbury
用戶:Prof. John E. Ladbury
研究方向:酪氨酸激酶介導的信號轉導
 
In my group we explore the structural, biophysical, and cellular outcomes of protein complex formation at membrane-bound receptors.
We use Microscale Thermophoresis (MST) in addition to other methods including isothermal titration calorimetry and surface plasmon resonance and find a good agreement between the methods. We are very pleased with the low material consumption, short measurement times and broad application range of MST.
(編者譯)
在我的研究小組,我們探索結構,生物物理和膜結合受體的蛋白質復合物。我們使用微量熱泳動儀(MST)發(fā)現(xiàn)該技術與其他傳統(tǒng)技術等溫滴定量熱法ITC和表面等離子體共振SPR之間的數據具有很好的吻合度,而且MST的樣品消耗量極低,測量時間很短,并且應用范圍廣泛。
比利時布魯塞爾自由大學
http://www.vib.be/en/research/scientists/Pages/Han-Remaut-Lab.asp
用戶:Prof. Han Remaut
研究方向:細菌細胞表面的結構分子生物學
 

 

 
We study the structural and molecular biology of bacterial adhesins and cell-surface filaments with respect to their function in bacterial pathogenesis, with the ultimate aim of developing a new generation of virulence-targeted antimicrobials.
MST really is opening up the easy determination of some Kd's that were hard to get to with other methods. So far, we measured protein-protein, protein-glycan, protein-small compound and protein-cofactor interactions with the Monolith NT.115.”
(編者譯)
我們研究細菌黏附素和細胞表面纖維絲的結構和分子生物學性質,用于闡述細菌致病機理,最終目標是開發(fā)新一代的有針對性的抗菌素。
MST真的是一種非常容易的方法用于測量Kd值,而且很多Kd值是難以使用其他傳統(tǒng)方法來完成的。到目前為止,我們使用標記款MST-NT.115測量了蛋白質-蛋白質,蛋白質-多糖,蛋白質-小化合物和蛋白-輔酶因子的相互作用。
美國辛辛那提大學醫(yī)學院
www.cincinnatichildrens.org/svc/find-professional/z/yi-zheng.htm
用戶:Yi Zheng, PhD
研究方向:信號轉導 –Rho GTPases
 
 
We are studying the molecular mechanisms of signal transduction processes involving Rho GTPases and are developing small molecule inhibitors and other strategies that interfere with specific Rho protein functions in leukemia, lymphoma and lung cancer.
We are very enthusiastic about MST as it quickly enabled us to measure protein:protein and protein:small molecule interactions that have been difficult to detect and quantify with standard biomolecule interaction technologies in the past years. MST provides us with a unique tool to validate the lead inhibitors that bind to specific sites of Rho GTPases or their regulators/effectors .
(編者譯)
我們正在研究Rho GTPases的信號轉導的分子機制的,并開發(fā)小分子抑制劑和其他方法用于特異性地干擾白血病、淋巴瘤、肺癌中的Rho蛋白質功能。
我們很喜歡MST技術,因為在過去的幾年里MST幫助我們快速完成測量蛋白質-蛋白質和蛋白質-小分子間的相互作用,很多時候很難使用其他標準的生物分子互作技術完成。MST為我們提供了一個獨特的工具來驗證重要的抑制劑是否結合到Rho GTPases的特異性位點或者結合到他的調節(jié)物/效應物。
 
德國慕尼黑大學Max-von-Pettenkofer研究所
www.mvp.uni-muenchen.de/virologie-ag2-forschung.html
用戶:Dr. Maria G. Vizoso Pinto
研究方向:病毒感染
 
“Our lab is developing diagnostic tools based on protein biochips and is studying the basic mechanisms underlying herpes virus infections” We are using MST for elucidating the function of viral proteins and its interaction with other viral and host cell proteins.
MST is simple to use and was quickly established in our lab. In particular we do appreciate the low instrument, consumables and maintenance costs associated with this new technology
(編者譯)
我們的實驗室正在開發(fā)基于蛋白質生物芯片的診斷工具,是研究皰疹病毒感染的基本機制”我們使用MST來闡明病毒蛋白的功能及其與其他病毒和宿主細胞蛋白質的相互作用。
MST使用簡單,很快就在我們的實驗室推廣使用開。我們特別欣賞新技術以及響應儀器的低耗材和維修成本。
丹麥奧爾胡斯大學
http://www.carb.dk/
用戶:Prof. Jens Stougaard
研究方向:多糖的功能
 
“We are interested in understanding the interactions between cells and organisms by investigating the role of polysaccharides exposed on cell surfaces and secreted polysaccharide signal molecules. We are applying MST to determine structural requirements for recognition of complex polysaccharides and the role of ligand-receptor interactions in the relationships between different cells and organisms.
MST is very useful for my lab since it allows to measure interactions in solution even in complex samples of membrane proteins. Also the small amount of sample material needed and the broad range of applications are very advantageous.”
(編者譯)
我們的實驗室通過研究暴露在細胞表面的多糖和分泌型多糖信號分子來揭示細胞與細胞以及有機體的相互作用。我們使用MST來測量復雜的多糖的結合以及配體-受體的結合以闡明不同細胞或機體間的相互作用。
MST在我們的實驗室非常有用,因為它可以測量膜蛋白在復雜的環(huán)境溶液中的相互作用。此外只需要極少的樣品消耗量和廣泛的應用范圍也是MST的優(yōu)勢。
德國柏林自由大學
http://www.physik.fu-berlin.de
用戶Prof. Dr. Joachim Heberle
研究方向膜蛋白的結構與功能
 
My lab focuses on the elucidation of the structure and function of membrane proteins. This topic includes the investigation of protein-protein and protein-substrate interactions. We are employing MST in binding studies of membrane sensors to their transducers as well as soluble transcription factors to DNA. We also plan for investigations of receptor – ligand interactions with this exciting new technique. MST turned out to be an extremely useful method in our lab to determine binding constants. It is advantageous that the fast and handy.
(編者譯)
我們的實驗室專注于研究膜蛋白的結構和功能,包括蛋白-蛋白以及蛋白-底物相互作用的研究。我們使用MST來研究膜蛋白感應器分子-傳感器分子-轉錄因子-DNA的結合反應。MST顯示出極好的優(yōu)點,包括快速和簡單測量,樣品消耗量很少等等。
德國雷根斯堡大學
www.uni-regensburg.de/laengst
用戶Prof. Dr. Gernot Längst
研究方向:染色質動力學和核組裝
 
Our lab focuses on the mechanisms and organization of DNA packaging inside the cell nucleus. We study the biochemical properties of molecular machines, the chromatin remodelers that regulate DNA accessibility and switch between ‘on’ and ‘off’ states of genes.
We are using MST for studying the activity of chromatin remodeling enzymes and to quantify their affinities towards DNA, RNA and other proteins.
MST is a big step forward towards quantitative biochemistry. MST is simple to use and was quickly established in our lab. It is a good alternative to the traditional electrophoretic mobility shift assays.
(編者譯)
我們的實驗室專注于研究細胞和之中的DNA包裝的機制,研究分子機器的生化特性,染色質重塑調節(jié)DNA組裝和調節(jié)基因的開關狀態(tài)。我們通過MST來研究染色質重組化酶并且定量他們與DNA,RNA以及其他蛋白之間的親和力。MST將定量生化技術推向一個新的臺階因為其簡單易學,并且非常好的取代了傳統(tǒng)的EMSA技術。
德國慕尼黑大學
www.molekularbiologie.abi.med.uni-muenchen.de
用戶Prof. Dr. Axel Imhof
研究方向:實驗胚胎學-組蛋白編碼
 
We are interested in the basic mechanisms in epigenetics which define and maintain the histone code and are using MST to measure the binding of “reader”-proteins to modified histone peptides as well as the activity and inhibition of “writer” enzymes including kinases, demethylases and methylases.
The new MST technology is easy to use and a good alternative to our standard enzymatic assays which are radioactive and measure endpoints only. It allows us to easily measure affinities for protein-peptide interactions in solution.
(編者譯)
我們對實驗胚胎學的定義和維護組蛋白準則的基本機制很感興趣,并且使用MST來測量“Reader”蛋白和修飾后的組蛋白多肽的結合以及“writer”酶(包括激酶、去甲基化酶、甲基化酶)的激活劑和抑制劑作用。
最新的MST技術非常簡單易學,是我們使用放射性終端測量等傳統(tǒng)方法的極好的取代技術,并且MST是在溶液之中測量蛋白-蛋白的相互作用的親和力。
德國法蘭克福大學
 
www.pzf.de/allg/research/schaefer.php
用戶Prof. Dr. med. Liliana Schaefer
研究方向炎癥和纖維化中的基質信號通路
 
The work of our group gave rise to the novel concept that under certain conditions matrix components may act as endogenous “danger” signals, which are recognized by innate immunity receptors, and are capable to trigger an inflammatory response reaction.
We are using MST to measure the interactions of endogenous and in vitro mutagenized proteoglycans of the extracellular matrix with their receptors and are very satisfied with the results obtained so far. The affinities are consistent with the biological readouts and confirm previous results gained with immunprecipitation and binding assays.
MST is a new technology we can definitely recommend for obtaining robust quantitative affinity data.”
(編者譯)
我們發(fā)現(xiàn)在特定化井下某些基質成分會變成內源性的“危險”信號,而這些異常的成分會被先天免疫受體識別,并且能夠引發(fā)炎癥反應。
我們使用MST來測量細胞外基質中的內源物/體內突變的蛋白聚糖與他們相關受體的反應,并且非常滿意目前獲得的所有數據,所得到的數據與其他生物方法預測的完全吻合并且驗證了之前使用免疫沉淀和結合試驗所獲得的數據。
MST是一種非常新的技術,我們推薦這個技術可以用來獲得非常好的定量的親和力數據。
德國慕尼黑大學基因中心
用戶Prof. Dr. Klaus Förstemann
研究方向:果蠅中miRNA的生物起源
 
We are studying the processing of microRNA precursors In Drosophila and are using MST to measure the binding of nucleolytic Enzymes and their specificity factors to RNA substrates.
The small amount of protein sample needed and the much faster measuring time compared with e.g. gel-shift assays are particular strengths of this new technique. It enables us to ask questions that we could not address before.
(編者譯)
我們正在研究果蠅前體細胞的MicroRNA發(fā)生過程,并且使用MST來測量核酶即特異性的因子結合到RNA上的反應。MST極少的蛋白樣品的消耗量和極短的測量時間使得他成為一個非常有價值的技術,它能幫助我們回答很多之前無法解釋的問題。
德國哥根廷大學物理化學系
用戶Prof. Dr. Andreas Janshoff
研究方向膜囊泡的作用
 
Membrane interaction and fusion plays a fundamental role in many cellular processes such as intracellular trafficking, fertilization, tissue formation or viral infection.
With the new MST technology we were able to measure the interaction of membrane vesicles, mediated by coiled coil-forming peptides. The technology requires only little sample material and is enabling for this type of experiments”(編者譯)
生物膜的融合以相互作用在細胞的多項過程中(細胞內運輸,受精,組織形成以及病毒感染)起到非常重要的基礎作用。通過MST技術,我們能夠測量螺旋狀多肽介導的囊泡的相互作用。該技術僅僅需要極少的樣品材料即可完成測量。
德國哥根廷大學有機化學與生物分子研究所
www.diederichsen.chemie.uni-goettingen.de/
用戶:Prof. Dr. Ulf Diederichsen
研究方向合成生物分子
 
We are interested in the biophysical properties of soluble and membrane bound biomolecules and are using MST to measure binding affinities and complex formation.
MST is straightforward and fits well in our technology portfolio to cover a broad spectrum of synthetic biomolecules including proteins, peptides and nucleic acids.
(編者譯)
生物膜的融合以相互作用在細胞的多項過程中(細胞內運輸,受精,組織形成以及病毒感染)起到非常重要的基礎作用。通過MST技術,我們能夠測量螺旋狀多肽介導的囊泡的相互作用。該技術僅僅需要極少的樣品材料即可完成測量。我們對游離的和膜上生物分子的生物物理學性質感興趣,并且使用MST來測量結合親和力和復合物的構象。
MST技術簡單易使用,很好的融入我們目前的技術體系,使我們能夠研究很大范圍的合成的生物分子,包括蛋白、多肽以及核酸。
麻省理工學院
 
 
http://web.mit.edu/lms/www/
用戶Dr. Shuguang Zhang張曙光教授
研究方向分子結構學
 
We are interested in molecular architectures especially to stabilize and analyze challenging smell receptors that belong to G protein-coupled receptors (GPCRs). GPCR is a super-family of receptors linked to many diseases. We want to measure the interactions of smell receptors with odorant that are very low molecular weight ligands. In the past that has been extremely difficult to detect and quantify other common molecular interaction technologies including SPR, and nearly impossible to use ITC for odorant-binding measurement because of the conformation and immobilization-sensitivities, or difficult and costly to obtain sufficient amount of receptors. The MST technology is a superb technology, it not only is label-free, hence reducing the measuring artifacts, but it also is surface-free that provides true measurement of finest molecular interactions. Thus it allows us much better analysis of the GPCR with their small-molecule ligands. We strongly recommend MST technology for its low material consumption (a few micrograms in a few microliters), short measurement times, inexpensive consumables, and broad application range in all molecule-interaction study.
我們的研究方向是分子結構學,尤其是G蛋白耦合受體(GPCRs)相關的嗅覺感受器分子穩(wěn)定性和分析。GPCR是與許多疾病相關的受體超級家族。我們想測量嗅覺感受器與香味劑分子間的相互作用,而香味劑分子通常都是非常低分子量的配體。在過去,因為香味劑結合反應的構象相關性和固定化敏感性,通過其他分子間相互作用的方法(包括SPR)一直很難檢測和量化的,并且由于需要昂貴和困難方法才能獲取微量的香味劑分子,通過ITC技術也是不可能的。MST技術是非常好的方法,它無需標記,還可以分析出實驗中的人為假象,而且不需要固定,能夠提供真實的分子間相互作用的數據,因此它可以讓我們更好地分析GPCR與小分子配體的作用。我們強烈推薦MST技術的幾個特點:極低的樣品消耗量(幾微克/幾微升),測量時間短,便宜的耗材,在分子相互作用研究中廣闊的應用范圍。
來源:Quantum量子科學儀器貿易(北京)有限公司
聯(lián)系電話:010-85120280(聯(lián)系我時,請說明是在生物器材網了解)
E-mail:info@qd-china.com

用戶名: 密碼: 匿名 快速注冊 忘記密碼
評論只代表網友觀點,不代表本站觀點。 請輸入驗證碼: 8795
Copyright(C) 1998-2025 生物器材網 電話:021-64166852;13621656896 E-mail:info@bio-equip.com
成人欧美一区二区三区黑人免费_亚洲永久无码7777KKK_国产欧美日韩综合精品一区二区_欧美在线一区二区三区
欧美人妻中文字幕在线| 99久久夜色精品| 欧美三级午夜理伦三级富婆| 久久精品国产亚洲av超碰| 亚洲精品一区二区三区影院| 精品久久久久久久| 毛片免费的| 《洗濯屋(H动漫)》中字| 国产一区二区三区在线观看视频 | 看久久毛片| 国产超级av在线| 高潮呻吟视频| 久久国产精品99久久人人澡| 国产手机精品一区二区| 久久精品欧美一区二区三区黑人| 国产乱码一区二区三区的解决方法| 国产欧美日韩视频在线| 日日干夜夜爽| 国产午夜精品一区二区三区四区 | 在线蜜臀av| 色婷婷国产| JAVAPARSER丰满白老师| xxoo亚洲精品久久久| 欧美精品中文字幕亚洲专区| 精品妇女一区二区三区| 色婷婷综合久久久久中文一区二区| 99久久国产精品免费热7788| а天堂中文在线官网在线| 国产一区二区精品久久| 97国产一区二区精品久久呦 | 日韩久久av| 国产精品女一区二区久久| 精品国产欧美一区二区| 午夜久久网| 成人免费在线小视频| 亚洲黄色一区二区三区| 亚洲综合精品熟女久久久40p| 久久久久久久亚洲| JAPANESE日本熟妇| 69人妻人人做人人爽人碰人人蜜臀 | 国产精品久久午夜伦鲁鲁|