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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。image.png

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


A key benefit of the biotin-streptavidin system is its ability to improve detection sensitivity. This, in large part, is due to the tetrameric conformation of streptavidin. One streptavidin protein can bind four biotin molecules with high affinity and selectivity. This multiplicity enables the amplification of weak signals and improves the detection sensitivity for medium- and low-abundance targets in mammalian cells or tissues with a simple workflow.

Another key benefit is the versatility of the biotin-streptavidin system. Because streptavidin can be conjugated to various reporter tags, it can be readily incorporated into virtually every immunoassay. For example, enzyme conjugates of streptavidin are widely used in enzyme-linked immunosorbent assays (ELISAs), while fluorescently labeled streptavidin, such as iFluor® 488 streptavidin, are widely used in cell surface labeling, fluorescence-activated cell sorting (FACS) and other fluorescence imaging applications.

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.
AAT Bioquest offers biotinylated secondary antibodies, proteins, nucleotides, and other small molecules for use in streptavidin-based amplification techniques. For assays in which a biotinylated probe is unavailable, we provide many biotinylation reagents and kits that enable researchers to chemically label proteins, nucleic acids, and surface materials to make custom biotinylated reagents.

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    

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