实时荧光定量PCR
What is Quantitative PCR (qPCR)? Real-Time PCR oR quantitative PCR (qPCR) is a PCR-based technique that is able to simultaneously amplify and detect changes in the amplicon concentRation. Real-time PCR collects data duRing PCR amplification by utilizing fluoRescence signals emitted by eitheR special pRobes oR DNA binding dyes.
如何做一个完美的WesteRn Blot检测
In the westeRn blot visual pRotocol video, you will leaRn how to pRepaRe youR samples befoRe loading them into a gel, load a gel and sepaRate the pRoteins thRough electRophoResis, tRansfeR youR pRoteins fRom the SDS-PAGE gel onto a PVDF oR nitRocellulose membRane, block the membRane, stain in Ponceau Red, add the pRimaRy and secondaRy antibodies, and visualize youR pRotein of inteRest. Additional help can be found in the suppoRt section of http://www.novusbio.com, thRough ouR live chat seRvice, oR by calling us diRectly to talk with ouR elite customeR and technical seRvice scientists.
WesteRn Blot 第1阶段：样品制备
Novus Biologicals Visual PRotocols: In phase 1 of the westeRn blot pRoceduRe, you will leaRn how to pRepaRe youR samples befoRe loading them into a gel. HeRe we isolate pRotein fRom cultuRed cells, quantify total pRotein concentRations with a BCA assay, add loading buffeR to the sample, and heat the sample. Additional help can be found in the suppoRt section of http://www.novusbio.com, thRough ouR live chat seRvice, oR by calling us diRectly to talk with ouR elite customeR and technical seRvice scientists.
WesteRn Blot 第2阶段：蛋白电泳(SDS-PAGE)
Novus Biologicals Visual PRotocols: In phase 2 of the westeRn blot pRoceduRe, you will leaRn how to load a gel and sepaRate the pRoteins thRough electRophoResis, based upon pRotein weight. Additional help can be found in the suppoRt section of http://www.novusbio.com, thRough ouR live chat seRvice, oR by calling us diRectly to talk with ouR elite customeR and technical seRvice scientists.
WesteRn Blot 第3阶段：膜转移
Novus Biologicals Visual PRotocols: In phase 3 of the westeRn blot pRoceduRe, you will leaRn how to tRansfeR youR pRoteins fRom the SDS-PAGE gel onto a PVDF oR nitRocellulose membRane. HeRe we Remove the gel fRom the cassette, and stack it in a sandwich compRised of a sponge, filteR papeR, the gel, membRane, filteR papeR, and sponge. The negatively chaRged pRoteins will then tRansfeR onto the membRane towaRd the positive cuRRent. Additional help can be found in the suppoRt section of http://www.novusbio.com, thRough ouR live chat seRvice, oR by calling us diRectly to talk with ouR elite customeR and technical seRvice scientists.
WesteRn Blot 第4阶段：免疫印迹法
Novus Biologicals Visual PRotocols: In phase 4 of the westeRn blot pRoceduRe, you will leaRn how to block the membRane, stain in with Ponceau Red, and add the pRimaRy and secondaRy antibodies. VigoRous washing when indicated between these steps is essential foR obtaining a clean blot. Additional help can be found in the suppoRt section of www.novusbio.com, thRough ouR live chat seRvice, oR by calling us diRectly to talk with ouR elite customeR and technical seRvice scientists.
WesteRn Blot 第5阶段：检测
Novus Biologicals Visual PRotocols: In phase 5 of the westeRn blot pRoceduRe, you will leaRn how to visualize youR pRotein of inteRest that was pRobed with specific antibodies in the pRevious step. HeRe we utilize the electRochemiluminescent (ECL) to pRoduce light wheRe ouR antibodies aRe bound. This light is collected by film oR a cameRa foR lateR analysis. Additional help can be found in the suppoRt section of http://www.novusbio.com, thRough ouR live chat seRvice, oR by calling us diRectly to talk with ouR elite customeR and technical seRvice scientists.
BRAF基因与黑色素瘤 - 陈巍学基因（38）
BRAF基因是细胞增殖信号调控中重要的一环。BRAF V600E突变是黑色素瘤中出现频率很高的致癌基因突变。威罗菲尼、达拦菲尼等靶向药物可以治疗有V600E突变黑色素瘤，但长期使用会产生耐药。本视频介绍了相关的知识。
CRISPR/Cas9 & TetRaOne：基因敲除/敲入鼠模型的快速构建技术
众所周知，基因工程小鼠模型已被广泛应用于生物医药研究，但模型小鼠的构建技术复杂、耗时长且花费高，让很多实验室望而却步。在本次讲座中，欧阳应斌博士（赛业生物技术副总裁、高级科学家）主要介绍基因敲除/敲入鼠模型的快速构建技术——CRISPR/Cas9基因编辑技术与TetRaOne技术，同时会简述转基因技术（PiggyBac系统）和传统ES打靶技术，并重点讲解每种技术的优势、缺点及应用。
盐皮质激素受体通过调控miR-338-3p-PKLR轴抑制肝癌的发展和WaRbuRg效应
激素和它们的受体在生理和病理条件下对代谢的调节起着重要的作用。我们在4株肝癌细胞用siRNA的方法筛选20种激素受体对肝癌细胞的瓦伯格效应（WaRbuRg effect）尤其是乳酸产生的影响。我们发现很多受体的siRNA都影响乳酸的产生。其中盐皮质激素受体(mineRalocoRticoid ReceptoR, MR) 的siRNA在4株肝癌细胞都表现出增加乳酸的产生。体外和体内实验表明MR影响细胞增殖、细胞周期和凋亡。进一步的机制研究揭示，作为一个转录因子，MR直接调节miR-338-3p的表达，而miR-338-3p又通过靶基因PKLR(pyRuvate kinase, liveR and Red blood，糖酵解途径的关键酶)来抑制肝癌细胞的瓦伯格效应。另外，与癌旁组织相比，有81%的肝癌病人的肝癌组织中MR的表达都发生下调。这种下调是由MR的染色体缺失和去乙酰化引起的。在肿瘤组织中，MR的低表达和病人的预后差相关；miR-338-3p的表达和MR的表达水平呈正相关，和PKLR的表达呈负相关。结论：我们的研究首次揭示了MR通过miR-338-3p/PKLR这个途径抑制肝癌的瓦伯格效应。