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AAT Bioquest，生物公司，品牌介绍AAT Bioquest是一家位于美国的生物公司，前身为ABD Bioquest，总部位于加利福尼亚州。专门从事光学检测技术十多年，一直致力于光谱学检测领域技术的创新和突破。其独特的光学检测技术，综合了化学、生物学和信息学等各个领域的研究，引领了比色、荧光和发光技术新一代光学探针的浪潮。AAT Bioquest在全球拥有强大的经验丰富的专业分销商网络，为从小型研究机构到《财富》500强企业的各类客户提供卓越的产品和定制服务。AAT Bioquest拥有超过5000项产品，包括Fluo-8®、Cal-520®、Cell Meter™、Amplite™、CytoTrace™和CytoTell™等产品线，这些品牌在生命科学研究领域获得了认可和好评。 AAT bioquest 提供酶、核酸、蛋白标记、小分子检测、细胞凋亡、钙离子、离子通道、膜电位检测等特色试剂盒及荧光探针。AAT Bioquest提供的客户定制服务包括比色、荧光和发光探针的合成，生化、细胞和诊断检测试剂的开发，以及使用AAT Bioquest已经验证的化合物库来筛选客户的目标化合物等。
l 酶的活性检测 l 核糖核酸检测 l 蛋白质化学 l 小分子检测 l 荧光标记探针 l 生物素及其衍生物 l 经典荧光标记染料 l 偶联剂 l 荧光淬灭剂 l 特优荧光标记染料 l 蛋白标记 Biotin-Streptavidin Conjugation System Illustration of the Streptavidin-Biotin interaction. The multivalent properties of streptavidin allow it to bind up to four biotin molecules with a high degree of affinity. Biotin is typically conjugated to an enzyme, antibody, or target protein. The widespread adoption of the biotin-streptavidin conjugation system is primarily due to two factors. The first is the relatively small size of the biotin and streptavidin molecules themselves, which allows for extensive binding to biologically active macromolecules, such as antibodies, without impedance to their functions (i.e., an antibody's binding site). The second is the specificity with which biotin and streptavidin bind to each other and the strength of the subsequent bond, which allows for powerful, stream-lined bioassay applications. Streptavidin tetramers have an extraordinarily high binding affinity for biotin with a dissociation constant (Kd) of approximately ~10-14 mol/L. This tight and specific binding is rapid and can withstand extremes in pH, temperature, organic solvents, and denaturing reagents. Advantages of the biotin-streptavidin system
Applications Enzyme-linked immunosorbent assay (ELISA) Immunohistochemistry (IHC) Power Styramide™ Signal Amplification, a superior replacement for tyramide Immunoblotting Immunofluorescence microscopy Cell surface labeling Affinity purification Fluorescence-activated cell sorting (FACS) Flow cytometry
Biotin Biotin is a small 244-dalton hapten molecule. Its high binding affinity for streptavidin is commonly exploited to detect and monitor biological targets of interest. Biotin exhibits two characteristics that make it ideal for bioconjugate development. First, biotin is relatively small in size. This permits labeling multiple biotin tags to a single protein or antibody without significantly impeding its biological reactivity. The second is the valeric acid side chain of the biotin molecule. This chain can be derivatized to facilitate the incorporation of various reactive groups used to conjugate biotin to proteins without altering biotin's binding affinity for streptavidin. Featured Products Fluorescent biotin derivatives Biotinylated secondaries Biotinylated nucleotides Biotinylated amino acids Reactive-biotin derivatives ReadiLink™ Protein Biotinylation Kit Amplite® Colorimetric Biotin Quantitation Kit
Fluorescent Biotin Derivatives Fluorescent biotin derivatives contain a fluorophore and a biotin moiety in the same molecule. These reagents are used for detecting and quantifying biotin-binding proteins by fluorescence. The strong quenching associated with streptavidin binding to fluorescent biotin can be used to precisely measure the concentration of streptavidin (or avidin). Product Ex/Em (nm) Ext. Coeff.1 FQY2 Unit Size Price Cat No. Fluorescein biotin 497/516 80,000 0.7900 5 mg $100 3017 Biotin-4-fluorescein *CAS 1032732-74-3* 492/518 80,000 0.7900 5 mg $100 3006 Cy5 biotin conjugate 650/669 250,000 0.27 5 mg $205 3100 Ext. Coeff. = molar extinction coefficient at their maximum absorption wavelength (Units = cm-1M-1). FQY = fluorescence quantum yield in aqueous buffer (pH 7.2). Biotinylated Secondary Antibodies Immunofluorescent stain of U+03b1-tubulin in HeLa cells. U+03b1-tubulin in fixed and permeabilized HeLa cells were labeled with rabbit anti-tubulin primary antibody, followed by incubation with biotinylated goat anti-rabbit IgG (H&L) (Cat No. 16794), and then visualized with iFluor® 555-streptavidin conjugate (Cat No. 16989). We offer biotinylated secondary antibodies to use with streptavidin-based amplification techniques. A key advantage to using a biotinylated secondary is the flexibility to use the same secondary antibody across multiple applications by using a different streptavidin conjugate. Readily available streptavidin conjugates HRP-streptavidin (imaging requires an HRP substrate) AP-streptavidin (imaging requires an AP substrate) iFluor®-streptavidin mFluor™-streptavidin APC, PE & tandem dye-streptavidins (The intense bright fluorescence and rapid photobleaching of phycobiliproteins are excellent for flow cytometry applications) Product Host/Isotype Species Reactivity Unit Size Price Cat No. Biotinylated goat anti-mouse IgG (H+L) Goat/IgG Mouse 1 mg $153 16729 Biotinylated goat anti-rabbit IgG (H+L) Goat/IgG Rabbit 1 mg $153 16794 Biotinylated Nucleotides Biotin-modified nucleoside triphosphate analogs, such as dUTP and dCTP, can be enzymatically incorporated into DNA or RNA fragments for use in fluorescence in situ hybridization (FISH), DNA arrays, microarrays, and other hybridization techniques. Standard enzymatic non-radioactive DNA labeling reactions include 3''-end labeling, cDNA labeling, nick translation, PCR, and random prime labeling. Each biotinylated nucleotide contains either an 11-, 14-, 16- or 20-atom spacer between the biotin and its attachment point on the nucleotide. This facilitates its detection and signal amplification by fluorophore and enzyme streptavidin conjugates or agarose and magnetic beads. Product Linker Length Unit Size Price Cat No. Biotin-11-dUTP 11 carbons 25 nmoles $100 17016 Biotin-16-dUTP 16 carbons 25 nmoles $100 17017 Biotin-20-dUTP 20 carbons 25 nmoles $100 17018 Biotin-14-dCTP 14 carbons 25 nmoles $100 17019 Biotin-11-dATP 11 carbons 25 nmoles $100 17014 Biotin-11-dGTP 11 carbons 25 nmoles $100 17015 Biotinylated Amino Acids Biotinylated peptides are suitable for immunodetection studies and biomedical screening assays that require immobilization onto streptavidin or avidin coated microtitre plates, beads, or membranes. Biotin can be linked to the N-terminus of peptides or the side chains of lysine (Lys), glutamic acid (Glu), or aspartic acid (Asp) during Fmoc solid phase synthesis. Incorporating a PEG-spacer in both FMOC-Glu(biotinyl-PEG)-OH and FMOC-Asp(biotinyl-PEG)-OH reduces steric hindrance between target peptide and streptavidin, leading to better biotin binding. The hydrophilic nature of the PEG-spacer minimizes non-specific interactions and improves the solubility of the reagent and biotinylated peptide. Product Labeling Principle Derivative Unit Size Price Cat No. FMOC-Lys(Biotin)-OH Uses FMOC-SPPS to prepare biotin-labeled peptides Lysine 100 mg $100 5020 FMOC-Glu(biotinyl-PEG)-OH Uses FMOC-SPPS to prepare biotin-labeled peptides Glutamic acid 100 mg $205 5026 FMOC-Glu(biotinyl-PEG)-OH Uses FMOC-SPPS to prepare biotin-labeled peptides Glutamic acid 1 g $998 5027 FMOC-Asp(biotinyl-PEG)-OH Uses FMOC-SPPS to prepare biotin-labeled peptides Aspartic acid 100 mg $205 5023 FMOC-Asp(biotinyl-PEG)-OH Uses FMOC-SPPS to prepare biotin-labeled peptides Aspartic acid 1 g $998 5024 Reactive-Biotin Derivatives Reactive biotin derivatives contain reactive moieties, such as succinimidyl esters or maleimides, that facilitate labeling biotin onto proteins, antibodies, and other macromolecules. When selecting a reactive biotin derivative, consider the following: Funtional group targeting - specific reactive moieties target and bind to their respective functional groups for targeted or non-selective biotinylation of proteins and other macromolecules (e.g., succinimidyl esters target primary amines and maleimides target thiol or sulfhydryl groups). Solubility - this influences biotinylation of the target protein or macromolecule. Spacer Arm Length (PEG) - enhance detection sensitivity of the target protein and reduces steric hindrance. ReadiView™ Biotinylation Reagents ReadiView™ biotin succinimidyl ester (Cat No. 3059) conjugated with specially designed color tag (CT) for easy determination of biotinylation. Determining a protein's degree of biotinylation is quite challenging because the intrinsic absorption of biotin is difficult to discern from proteins and nucleic acids. To facilitate this determination, we offer a series of chromophoric reactive biotin derivatives, ReadiView™ biotin. These novel biotinylation reagents contain a specially designed 'Color Tag (CT)' optimally positioned between two spacer arms. These spacer arms reduce steric hindrance and improve water solubility, while the CT tag makes the degree of biotinylation readily quantifiable by calculating the absorption ratio of 280 nm/385 nm. The CT tag does not affect biotins complexation with streptavidin and has a minimal quenching effect on fluorescent streptavidin conjugates. ReadiView™ biotin is available in amine-reactive, thiol-reactive, and click-chemistry formats. Product Chemical Reactivity Reactive Moiety Unit Size Cat No. Biotin, succinimidyl ester *CAS 35013-72-0* Amine Succinimidyl Ester 100 mg 3002 Biotin-X, succinimidyl ester *CAS 72040-63-2* Amine Succinimidyl Ester 25 mg 3010 Biotin PEG2 succinimidyl ester Amine Succinimidyl Ester 25 mg 3016 Biotin PEG4 succinimidyl ester Amine Succinimidyl Ester 25 mg 3022 Biotin C2 maleimide Thiol Maleimide 25 mg 3005 Biotin PEG2 maleimide *CAS 305372-39-8* Thiol Maleimide 5 mg 3015 Biotin PEG2 amine *CAS 138529-46-1* Aldehyde, Carboxylic Acid, Ketone Amine 5 mg 3014 Biotin PEG3 amine Aldehyde, Carboxylic Acid, Ketone Amine 5 mg 3024 Biotin cadaverine Aldehyde, Carboxylic Acid, Ketone Cadaverine 5 mg 3004 Biotin ethylenediamine *CAS 216299-38-6* Aldehyde, Carboxylic Acid, Ketone Ethylenediamine 5 mg 3003 Product Chemical Reactivity Unit Size Price Cat No. ReadiView™ biotin succinimidyl ester Amines Pegylated 5 mg $106 3059 ReadiView™ biotin maleimide Thiol or sulfhydryl groups Pegylated 5 mg $217 3058 ReadiView™ biotin hydrazide Aldehyde, Carboxylic acid, Ketone Pegylated 5 mg $217 3055 ReadiView™ biotin amine Carbonyl Pegylated 5 mg $217 3053 ReadiView™ biotin acid Amine Pegylated 5 mg $106 3050 Enzyme-Labeled Streptavidin Conjugates IHC detection of EpCAM in FFPE lung adenocarcinoma tissue. Human adenocarcinoma tissue sections were incubated with rabbit anti-EpCAM primary antibody followed by biotinylated goat anti-rabbit IgG (H&L). The samples were then incubated with HRP-streptavidin, and the signal was developed using ReadiUse™ Stayright™ Purple. Enzyme-labeled streptavidin conjugates are commonly used in enzyme-linked immunosorbent assay (ELISA), immunohistochemistry, immunoblotting, and in situ hybridization techniques. Enzymes horseradish peroxidase (HRP) and alkaline phosphatase (ALP) are extensively used as reporter tags because of their high turnover rate, stability, ease of conjugation, and low cost. To maximize retention of both enzyme and protein activity, our HRP-streptavidin and AP-streptavidin conjugates are prepared at a 1:1 (enzyme to streptavidin) labeling ratio. Chromogenic, fluorogenic, and chemiluminescent substrates are available for these enzyme conjugates. Product Enzyme Unit Size Cat No. HRP-streptavidin conjugate Horseradish peroxidase (HRP) 1 mg 16920 AP-streptavidin conjugate Alkaline phosphatase (AP) 1 mg 16921 Product Detection Abs/Ex (nm) Ex (nm) Unit Size Cat No. ReadiUse™ ABTS Chromogenic 420 1 L 11001 ReadiUse™ TMB Chromogenic 650 100 mL 11012 ReadiUse™ TMB Chromogenic 650 1 L 11003 Amplite® ADHP Fluorogenic 571 584 25 mg 11000 Amplite® Blue Fluorogenic 324 409 25 mg 11005 Amplite® Red Fluorogenic 571 584 1000 Assays 11011 Amplite® IR Fluorogenic 648 668 1 mg 11009 Luminol Chemiluminescent 355 412 1 g 11050 Product Detection Abs/Ex (nm) Ex (nm) Unit Size Cat No. pNPP *CAS 4264-83-9* Chromogenic 405 25 mg 11619 FDP *CAS 217305-49-2* Fluorogenic 498 517 5 mg 11600 MUP, disodium salt *CAS 22919-26-2* Fluorogenic 360 448 25 mg 11610 MUP, disodium salt *CAS 22919-26-2* Fluorogenic 360 448 10 g 11612 DiFMUP Fluorogenic 358 450 5 mg 11627 SunRed™ Phosphate Fluorogenic 653 661 5 mg 11629 D-Luciferin phosphate *CAS 145643-12-3 Chemiluminescent 5 mg 12512 Fluorescent Streptavidin Conjugates Immunofluorescent stain of &alpha-tubulin in HeLa cells. &alpha-tubulin in fixed and permeabilized HeLa cells were incubated with rabbit anti-tubulin primary antibody, followed by incubation with biotinylated goat anti-rabbit IgG (H&L) (Cat No. 16794), and then visualized with iFluor® 647-streptavidin conjugate (Cat No. 16966. Nuclei were counterstained using DAPI (Cat No. 17507.) Fluorescent streptavidin conjugates are widely used in fluorescence imaging to detect biotinylated antibodies, ligands and DNA probes for in situ hybridization techniques, immunohistochemistry and multicolor flow cytometry. We offer a comprehensive portfolio of fluorescent streptavidin labeled with iFluor® dyes, mFluor™ dyes, phycobiliproteins, tandem dyes, and more. Benefits of our iFluor® streptavidin conjugates include exceptionally bright fluorescence, excellent photostability, and pH insensitive fluorescence over a wide molar range. iFluor® Streptavidin Conjugates Streptavidin conjugates labeled with iFluor® dyes exhibit exceptionally bright and photostable fluorescence, outperforming Alexa Fluor® and other spectrally similar conjugates. iFluor® streptavidin conjugates are designed for various cell analysis and protein analysis applications, including imaging, flow cytometry, and fluorescent western blotting. As a precaution, avoid using blue fluorescent streptavidin conjugates when detecting low abundance targets. Blue fluorescent dyes have lower fluorescence and can experience a higher non-specific background than other dyes. Product Ex/Em (nm) Filter Set Ext. Coeff.1 FQY2 Unit Size Price Cat No. iFluor® 350 Streptavidin 344/448 DAPI 20,000 0.95 1 mg $162 16980 iFluor® 405 Streptavidin 402/425 DAPI 37,000 0.91 1 mg $162 16982 iFluor® 488 Streptavidin 491/516 FITC 75,000 0.9 1 mg $162 16985 iFluor® 514 Streptavidin 527/554 TRITC 80,000 0.83 1 mg $162 16986 iFluor® 532 Streptavidin 543/563 TRITC 90,000 0.68 1 mg $162 16987 iFluor® 546 Streptavidin 541/557 TRITC 100,000 0.67 200 μg $84 16958 iFluor® 555 Streptavidin 556/569 TRITC 100,000 0.64 1 mg $162 16989 iFluor® 568 Streptavidin 568/587 TRITC 100,000 0.57 200 μg $84 16960 iFluor® 594 Streptavidin 587/603 Texas Red 180,000 0.53 1 mg $162 16992 iFluor® 633 Streptavidin 638/652 Texas Red 250,000 0.29 1 mg $162 16995 Ext. Coeff. = Extinction coefficient at their maximum absorption wavelength. The units of extinction coefficient are cm-1M-1. FQY = fluorescence quantum yield in aqueous buffer (pH 7.2). N/D = Not determined. mFluor™ Streptavidin Conjugates for Flow Cytometry Flow cytometry analysis of HL-60 cells stained with (Red) or without (Green) 1ug/ml Anti-Human HLA-ABC-Biotin and then followed by mFluor™ Blue 570-streptavidin conjugate (Cat No.16935). Streptavidin conjugates labeled with mFluor™ dyes exhibit minimal self-quenching, producing intensely fluorescent and photostable conjugates for flow cytometry. mFluor™ streptavidin conjugates are designed for optimal excitation by one of the major laser lines commonly equipped in flow cytometers, such as the 405 nm, 488 nm, 532 nm, 561 nm, or 633 nm laser lines. Streptavidin Conjugates Ex (nm) Em (nm) Filter Set Cat No. mFluor™ Violet 450 Streptavidin 405 444 450/40 16930 mFluor™ Violet 510 Streptavidin 408 503 525/50 16931 mFluor™ Violet 540 Streptavidin 393 536 525/50 16932 mFluor™ Blue 570 Streptavidin 505 564 575/26 16935 mFluor™ Green 620 Streptavidin 522 621 610/20 16938 mFluor™ Yellow 630 Streptavidin 609 626 610/20 16942 mFluor™ Red 700 Streptavidin 685 704 730/45 16946 mFluor™ Red 780 Streptavidin 629 780 780/60 16948 Streptavidin APC, PE, PerCP and Tandem Dye Conjugates for Flow Cytometry Streptavidin PE, APC, and PerCP conjugates are commonly used in applications that require high sensitivity but not photostability, primarily flow cytometry, FACS, immunophenotyping, and microarrays. The unusually bright fluorescence of phycobiliproteins is due to their unusually high extinction coefficients and quantum yields. Tandem dye conjugates compromise of a donor phycobiliprotein (e.g., APC or PE) labeled to acceptor iFluor® dyes that emit fluorescence at a longer wavelength. Using fluorescence resonance energy transfer (FRET), light absorbed by the donor phycobiliprotein results in fluorescence emission of the acceptor. In flow cytometry, tandem dyes are ideally suited for multicolor parametric analysis of cells due to their exploitation of a single excitation source and their significantly large Stokes shifts. Streptavidin Conjugates RPE Streptavidin 564 574 575/26 16900 APC Streptavidin 651 660 660/20 16902 PerCP Streptavidin 477 678 695/40 16905 RPE-iFluor® 647 Streptavidin 568 666 670/14 16906 RPE-iFluor® 750 Streptavidin 566 778 780/60 16907 APC-iFluor® 750 Streptavidin 651 791 780/60 16908 RPE-Cy7 Streptavidin 566 778 780/60 16916 Streptavidin-Xtra™ Conjugates Comparison of HeLa cells stained with Streptavidin-Xtra™ IF488 and Streptavidin-Alexa Fluor® 488. HeLa cells were fixed with 4% paraformaldehyde for 30 minutes, permeabilized with 0.02% Triton™ X-100 for 10 minutes, blocked with 1% BSA for 1 hour and labeled with 1 μg/mL U+03b1-tubulin Mouse mAb for 1 hour at room temperature. Biotinylated goat anti-mouse IgG (H+L) (Cat No. 16729) was used for detection of U+03b1-tubulin and visualized with Streptavidin-Xtra™ iFluor 488 (Cat No. 46001) and Streptavidin-Alexa Fluor® 488. Cells were counterstained with Hoechst 33342 (Cat No. 17535). The optimal degree of labeling of Streptavidin-Xtra™ iFluor® conjugates produce the brightest fluorescent streptavidin conjugates for the detection of biotinylated antibodies in cell imaging, flow cytometry, and western blot applications. Streptavidin-Xtra™ iFluor® conjugates image low-abundance targets with greater sensitivity and detail than traditional streptavidin Alexa Fluor® conjugates. Advantages Bright and photostable fluorescence without significant self-quenching High signal-to-noise ratios: 3~5 fold signal improvement versus Alexa Fluor® pH insensitive fluorescence over a wide molar range Streptavidin-Xtra™ iFluor® 488 491 516 FITC 46001 Streptavidin-Xtra™ iFluor® 555 556 569 TRITC 46003 Streptavidin-Xtra™ iFluor® 594 587 603 Texas Red 46005 Streptavidin-Xtra™ iFluor® 647 654 669 Cy5 46007 Cat# Product Name 3001 Biotin *CAS 58-85-5* 1 g 79 3002 Biotin, succinimidyl ester *CAS 35013-72-0* 100 mg 205 3003 Biotin ethylenediamine *CAS 216299-38-6* 10 mg 100 3004 Biotin cadaverine 25 mg 100 3005 Biotin C2 maleimide 25 mg 310 3006 Biotin-4-fluorescein *CAS 1032732-74-3* 5 mg 100 492 518 3007 Biotin hydrazide *CAS 66640-86-6* 25 mg 79 3009 Biotin-X NTA [Biotin-X nitrilotriacetic acid, potassium salt] *CAS 856661-92-2* 1 mg 100 3010 Biotin-X, succinimidyl ester *CAS 72040-63-2* 25 mg 79 3014 Biotin PEG2 amine *CAS 138529-46-1* 5 mg 100 Cat# Product Name 16900 RPE-streptavidin conjugate 100 ug 100 16901 RPE-streptavidin conjugate 1 mg 310 16902 APC-streptavidin conjugate 100 ug 153 16905 PerCP-streptavidin conjugate 100 ug 205 16906 RPE-iFluor® 647-streptavidin conjugate 100 ug 205 16907 RPE-iFluor® 750-streptavidin conjugate 100 ug 205 16908 APC-iFluor® 750-streptavidin conjugate 100 ug 205 16910 FITC-streptavidin conjugate 1 mg 153 16911 Texas Red®-streptavidin conjugate 1 mg 153 16912 Cy3®-streptavidin conjugate 1 mg 153 |
