Biotin
mRNA synthesis
In vitro transcription of capped mRNA with modified nucleotides and Poly(A) tail
Tyramide Signal Amplification (TSA)
TSA (Tyramide Signal Amplification), used for signal amplification of ISH, IHC and IC etc.
Phos Binding Reagent Acrylamide
Separation of phosphorylated and non-phosphorylated proteins without phospho-specific antibody
Cell Counting Kit-8 (CCK-8)
A convenient and sensitive way for cell proliferation assay and cytotoxicity assay
SYBR Safe DNA Gel Stain
Safe and sensitive stain for visualization of DNA or RNA in agarose or acrylamide gels.
Inhibitor Cocktails
Protect the integrity of proteins from multiple proteases and phosphatases for different applications.
NHS-生物素修饰是一种特定的赖氨酸探针,可以耦合质谱检测,在过去几年中越来越多地被应用,可以评估蛋白质或复合物的氨基酸可及性,作为可替代的行之有效的方法1。
许多应用已经报道NHS-biotin作为特定的赖氨酸探针2,其应用越来越多,已经有大量的报道阐述了其应用2。可以用于判定氨基酸的表面可及性,用于表位作图中划定蛋白质-蛋白质界面4。
Sulfo-NHS-biotin试剂与模型肽33和3,30-Dithiobis[sulfosuccinimidyl propionate] (DTSSP) 发生反应,标记含有羟基的氨基酸5。如果含羟基的基团位于组氨酸残基的两个位置,丝氨酸、酪氨酸和苏氨酸残基可以与Sulfo- NHS-biotin 发生反应6,这些羟基通过氢键与组氨酰残基的相互作用,增强这些含羟基基团的内在反应性,有助于增加其亲核性。
使用NHS-生物素衍生物检测DTSSP的开拓观察:丝氨酸和酪氨酸残基的羟基基团也与伯胺试剂反应,丝氨酸标签可能会丢失,导致脱水肽形成。生物素试剂的应用导致通过链霉亲和素的亲和富集3,通过分光光度测定法对每个多肽的标签进行绝对定量。
使用整套Sulfo-NHS-biotin试剂可以检测到不同的离子的有效且可靠的离子分配。
参考文献:
1. G. Gabant, J. Augier et al, Assessment of solvent residues accessibility using three Sulfo-NHS-biotin reagents in parallel: application to footprint changes of a methyltransferase upon binding its substrate, J. Mass Spectrom. 2008; 43: 360–370
2. Glocker MO, Borchers C, Fiedler W, Suckau D, Przybylski M. Molecular characterization of surface topology in protein tertiary structures by amino-acylation and mass spectrometric peptide mapping. Bioconjugate Chem. 1994; 5: 583.
3. Azim-Zadeh O, Hillebrecht A, Linne U, Marahiel MA, Klebe G, Lingelbach K, Nyalwidhe J. Use of biotin derivatives to probe conformational changes in proteins. Journal of Biological Chemistry 2007; 282: 21609
4. Borch J, Jorgensen TJ, Roepstorff P. Mass spectrometric analysis of protein interactions. Current Opinion in Chemical Biology2005; 9: 509.
5. Swaim CL, Smith JB, Smith DL. Unexpected products from the reaction of the synthetic cross-linker 3, 3’ dithiobis(sulfosuccinimidyl propionate), DTSSP with peptides. Journal of the American Society for Mass Spectrometry 2004; 15:736.
6. Miller BT, Kurosky A. Elevated intrinsic reactivity of seryl hydroxyl groups within the linear peptide triads His-Xaa-Ser or Ser-Xaa-His. Biochemical and Biophysical Research Communications 1993; 196: 461.
Physical Appearance | A solid |
Storage | Store at -20°C |
M.Wt | 244.31 |
Cas No. | 58-85-5 |
Formula | C10H16N2O3S |
Synonyms | D-Biotin,Vitamin H |
Solubility | ≥24.4 mg/mL in DMSO; insoluble in H2O; insoluble in EtOH |
Chemical Name | 5-[(3aS,4S,6aR)-2-oxo-1,3,3a,4,6,6a-hexahydrothieno[3,4-d]imidazol-4-yl]pentanoic acid |
SDF | Download SDF |
Canonical SMILES | C1C2C(C(S1)CCCCC(=O)O)NC(=O)N2 |
运输条件 | 蓝冰运输或根据您的需求运输。 |
一般建议 | 不同厂家不同批次产品溶解度各有差异,仅做参考。若实验所需浓度过大至产品溶解极限,请添加助溶剂助溶或自行调整浓度。溶液形式一般不宜长期储存,请尽快用完。 |
生物素化方法[1]: | |
样品 |
蛋白 |
溶解方法 |
可以溶于有机溶剂DMSO或者DMF中,为了获得更高的浓度,可以将离心管在37℃加热10分钟和/或在超声波浴中震荡一段时间。原液可以在-20℃以下储存几个月。 |
反应时间 |
室温下放置1h |
应用 |
生物素标记蛋白: Biotinyl-N-hydroxysuccinimide (BNHS) 能够诱导生物素部分进入蛋白,为了得到期望的BNHS与蛋白游离氨基的摩尔比值,将0.1 M NaHCO(含10 mg/ml蛋白质)与不同体积的0.1 M 的BNHS溶液混合,混合物室温下孵育1h后,在4℃下用不同浓度的PBS透析24h。透析之后,加入等体积的甘油后-20℃保存。BNHS 溶液应现配现用。2, 4, 6-三硝基苯磺酸可以用来检测对照或者生物素化的蛋白质中的游离氨基。与IgG共价结合的总biotin可以通过链霉素结合试验进行检测。 |
References: [1]. JEAN-LUC GUESDON, THERESE TERNYNCK AND STRATIS AVRAMEAS. The Use of Avidin-Biotin Interaction in Immunoenzymatic Techniques. THE JOURNAL OF HISTOCHEMISTRY AND CYTOCHEMISTRY. 1979. |
质量控制和MSDS
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