| Molecular Weight: | 212.25 |
| Formula: | C13H12N2O |
| Purity: | ≥98% |
| CAS#: | 442-51-3 |
| Solubility: | DMSO up to 100 mM |
| Chemical Name: | 7-methoxy-1-methyl-9H-pyrido[3,4-b]indole |
| Storage: | Powder:4oC 1 year. DMSO:4oC3 month;-20oC 1 year. |
Biological Activity:Harmine is a potent, selective and orally bioavailable DYRK1A inhibitor with IC50 of 80 nM. It inhibits phosphorylation of tau directly by DYRK1A (IC50 ~700 nM). It has >10-fold selectivity over DYRK3 and DYRK2 (IC50 ~800 nM and 900 nM respectively. Harmine is also a unique regulator of PPARγ expression that acts by inhibiting the Wnt signalling pathway in a cell-specific manner. It attenuates inflammatory gene expression (TNFα, IL-1β, iNOS) and macrophage accumulation in adipose tissue. Administration of harmine (30 mg/kg) to obese db/db mice resulted in reduced blood glucose, free fatty acids, and triglyceride levels, delayed hyperglycemia, and improved insulin sensitivity. Being function as a new class of human beta cell mitogenic compound, by using three different mouse and human islet in vivo-based models, harmine is able to induce beta cell proliferation, increase islet mass and improve glycemic control. The nuclear factors of activated T cells (NFAT) family of transcription factors are defined as likely mediators of human beta cell proliferation and differentiation.How to Use:In vitro: Harmine was used at 10 µM final concentration in various in vitro assays.In vivo: Harmine was dosed orally to obese db/db mice at 30 mg/kg. Harmine was dosed by intraperitoneal injection at 10 mg/kg in mouse partial pancreatectomy model (PPX), Euglycemic human islet transplantation model and diabetic marginal mass human islet transplantation model.Reference:
- 1.Bain J, et al. The selectivity of protein kinase inhibitors: a further update. (2007) Biochem J. 408(3):297-315.
- 2.Waki H, et al. The small molecule harmine is an antidiabetic cell-type-specific regulator of PPARgamma expression. (2007) Cell Metab. 5(5):357-70.
- 3.Egusa H, et al. The small molecule harmine regulates NFATc1 and Id2 expression in osteoclast progenitor cells. (2011) Bone. 49(2):264-74.
- 4.Smith B, et al. Recent advances in the design, synthesis, and biological evaluation of selective DYRK1A inhibitors: a new avenue for a disease modifying treatment of Alzheimer's? (2012) ACS Chem Neurosci. 3(11):857-72.
- 5.Wang P, at al. A high-throughput chemical screen reveals that harmine-mediated inhibition of DYRK1A increases human pancreatic beta cell replication. (2015) Nat Med. In press.
Harmine_spec.pdf
Harmine_MSDS.pdfProducts are for research use only. Not for human use.ebiomall.com
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、
Cell-Based ELISA
的优点:
Cell-Based ELISA
(基于细胞的
ELISA
)是一种全新的
ELISA
技术,有两个最突出的优点:
1.1
不需要抽提蛋白、包被微孔板:细胞直接在微孔板里培养,待检测的时候,将细胞固定在微孔板上并
进行通透处理即可。这样就避免了抽提蛋白时,由于客观和主观上引起样品的损失而导致实验结果在一定
程度上偏离了实际情况。
同时不用包被微孔板,
简化了实验流程,
有助于提高效率。
科研人员在一个
96
孔
酶联板上,便能检测目标细胞蛋白经刺激或抑制作用后的表现。由于省去抽提蛋白和裂解细胞的步骤,样
本的损失也能降到最低,比起其他普通的
ELISA
测定方法,这项全新的
ELISA
技术能更快速、更方便
地一次检测大量的细胞内蛋白。
1.2
可同时检测两种不同蛋白:
封闭后加入两种抗不同蛋白且来源于不同宿主的一抗,
然后再加入不同的
二抗,加入两种荧光底物,检测两个波长。同时检测两种蛋白的好处是显而易见的,可以减少工作量,此
外还可以满足一些特殊的实验需要,例如,需要测定某个蛋白的磷酸化比例,就需要测定磷酸化蛋白的数
量和总蛋白的数量,这两个测定在同一次实验进行,有助于消除实验误差,得到更为精确的实验结果。
2
、
Cell-Based ELISA
的两种技术:
某些公司发展了双通道
Cell-Based ELISA
技术;
双通道与单通道
Cell-based ELISA
比较:
双通道
cell-based ELISA
,顾名思义,即一次可以同时检测两种目的蛋白,
R&D Systems
提供的
Cell-based ELISA
就是双通道
cell-based ELISA
,原理略:(需要荧光检测方式和相应仪器);
单通道
cell-based ELISA
,即一次只能检测一种目的蛋白,他和普通
ELISA
的主要区别在于样品处理过
程
3
、
cell-based ELISA
的应用:
该产品,最多发展起来的是用于检测磷酸化和非磷酸化蛋白的相对含量;
4
、有
cell-based ELISA
产品的公司:目前,多家
elisa
产品提供
商均
提供
cell
based
elisa
试剂盒,
如
Rnd
systems
,
raybiotech
,
ebioscience
,
millipore
等公司;
5
、如何自己进行
cell based elisa
实验?
事实上,根据单通道的
cell based elisa
原理,可以自己建立
cell based elisa
实验系统;
细胞加入培养板中;
加入刺激物或者抑制剂进行培养;
对细胞进行固定或者封闭;
加入第一抗体;
加入
HRP
结合的二抗(二抗的选择,同
western blot
);
加入底物进行显色;
我做的是细胞因子的刺激和抑制某条通路后观察是否有影响,分组为空白组,空白+抑制剂,刺激组,刺激+抑制剂,最开始用的单因素方差分析,LSD-T和SNK-Q检验,但是同学说我这里面有两个处理因素,所以不能单因素方差分析,应该直接空白和空白+抑制,空白和刺激,刺激和刺激+抑制剂进行独立样本T检验,现在脑子是混乱的,拜托园子里的大神们帮我看看,感激不尽!!
支原体培养则是取样后在培养基上培养,看有多少支原体菌落会长出,是比较直观和可信的结果。
总体来讲,这两种检查手段可信度都较高,结合一起,不仅可以可靠的知道有无解脲支原体感染,还能知道感染是否严重。
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MaggotsFasterThanScalpelinWoundDebridement
December19,2011—Maggotdebridementtherapy(MDT)appearstobemoreeffectiveforwounddebridementcomparedwithconventionaltherapy,butonlyat1week;afterthattime,anothertypeofdressingshouldbeused,newresearchsuggests.
KristinaOpletalovà,MD,fromtheDepartmentofDermatology,UniversityofCaen,France,andcolleaguespublishedonlineDecember19intheArchivesofDermatology.
MedicalmaggotswereapprovedbytheUSFoodandDrugAdmiNISTrationasamedicaldeviceforwounddebridementin2004.Accordingtotheresearchers,useofmaggotsintreatingwoundsisassociatedwitheffectivewounddebridement,antibacterialeffects,andstimulationofwoundhealing.
However,theypointout,"[r]elativelyfewclinicalstudieshavebeenconductedandtheresultsarenotclear,partlyowingtomethodologicassessmentproblems."
InthecurrentProspective,randomizedcontrolled,phase3clinicaltrial,theresearcherssoughttodeterminetheefficacyofbaggedlarvaeonwounddebridementincomparisonwithconventionaltreatment.
TheprimaryobjectivewastocomparethemeanpercentageofsloughinwoundstreatedwithMDTwiththatofconventionaltreatmentatday15.Thestudyincluded119patientswithanonhealing,sloughywoundthatwas40cm2orsmallerandlessthan2cmdeep.Patientsalsohadananklebrachialindexof0.8orhigher.
Treatmentwasadministeredduringa2-weekhospitalstay.Conventionaltreatmentconsistedofsurgicaldebridement3timesaweekwithascalpel,withuseoftopicalanesthesia.TheMDTwasadministeredusinganencloseddressing(Vitapad,BioMondeLaboratories)containing80sterilemaggots.Atdischarge,aconventionaldressingwasapplied,andpatientswerefollowed-upatday30.
DebridementbyMDTwassignificantlyfasterthansurgicaldebridementduringthefirstweekoftreatment,reachingthesamelevelthecontrolgroupreachedatday15.NobenefitforMDTcomparedwithconventionaltreatmentinhealingrateswasobserved.Atday8,54.5%intheMDTgroupvs66.5%inthecontrolgroup(P=.04)hadevidenceofsloughandwoundhealing.However,byday15,themeanpercentageofsloughwas55.4%intheMDTgroupand53.8%inthecontrolgroup(P=.78).
"AthoughMDTshowsnosignificantbenefitatday15comparedwithconventionaltreatment,debridementbyMDTissignificantlyfasterandoccursduringthefirstweekoftreatment,"theresearchersconclude."Becausethereisnobenefitincontinuingthetreatmentafter1week,anothertypeofdressingshouldbeusedafter2or3applicationsofMDT."
Painscoresweresimilarandmildinbothgroups,althoughincontrasttoconventionaltreatment,MDTwasperformedwithouttopicalanesthesia.
Accordingtotheresearchers,noneofthepatientswerereticentaboutundergoingMDT."[A]crawlingsensationonthewoundwasrarelyandalmostequallynotedinbothgroups,revealingthatthesensationwassubjective,"Dr.Opletalovàandcolleaguespointout.
TwoquestionsregardingMDTremainunanswered,theauthorsnote."Candebridementbeimprovedusingmoremaggotsperdressing?Ifso,wouldthesedressingsbemorepainful?Furtherstudiesareneededtoanswerthesequestions."
ThestudywassupportedbygrantsfromtheClinicalResearchHospitalProgramandfromtheFrenchSocietyofDermatology.Theauthorshavedisclosednorelevantfinancialrelationships.

