The conjugate binds four molecules of biotin with high affinity k d 1015 m1. A 60kda extracellular protein of streptomyces avidinii with four high affinity biotin binding sites. Streptavidin polyhrp conjugate is designed to deliver the highest sensitivity and low background in immunoassays where sample volume is limited or when the target molecule is present at low levels. Thermo scientific pierce high sensitivity streptavidin.
Streptavidinhrp conjugates have a very high turnover rate, coupling high sensitivity with short. In the preparation of this product, invitrogen uses highly purified streptavidin and. This poly hrp conjugate is designed to deliver the highest sensitivity and low background in immunoassays where sample volume is limited or when the target molecule is present at low levels. Streptavidin hrp is useful for detecting biotinylated antibodies, e. Streptavidin hrp conjugate elisa grade immunoreagents. Thermo scientific pierce high sensitivity streptavidin hrp, 0. Thermo scientific pierce high sensitivity streptavidinhrp, prediluted amplify signal in western blot, elisa or other biotinbased detection methods with this exclusive, high signaltonoise, peroxidaseconjugated, readytouse streptavidin. Streptavidin has a molecular weight of 60 kd and has 4 subunits. Streptavidin poly hrp conjugate is designed to deliver the highest sensitivity and low background in immunoassays where sample volume is limited or when the target molecule is present at low levels.
Streptavidin hrp m00091 is coupled steptavidin with horseradish peroxidase hrp. Hrpstreptavidin conjugate comprises streptavidin as the biotinbinding protein with hrp covalently attached as the enzyme label. Im currently using a streptavidinhrp conjugate along with a primary and biotinlabeled secondary to detect proteins in whole cell lysates. Such claim would disregard the fact that well developed elisa kit is an integrate, finely balanced system. Streptavidin is a tetrameric bacterial protein isolated from streptomyces avidinii providing 4 highaffinity biotin binding sites figure 1. Avidin and streptavidin reagents are powerful tools. It does not guarantee that antibodyconjugate combinations have been tested for use in the listed applications.
Streptavidinbinding peptide sbptagged smc2 allows single phosphataseconjugated antibody jackson immunoresearch laboratory for 1 hour at 1. The hrpluminolh2o2 chemiluminescent system with high sensitivity was chosen as the detection system. Product data sheet pierce high sensitivity streptavidinhrp. These conjugates bind to biotin labeled proteins in western. Then the signal amplification was achieved through the interaction between the biotinylated antitsh mab and the hrpstreptavidin conjugate. Whether detecting your signal by fluorescent, colorimetric or chemiluminescent methods, these plates make it easy to develop an elisa for any sample with a low.
These conjugates are typically used as secondary reagents to detect biotinylated probes such as primary antibodies for flow cytometry, western blotting, immunofluorescence staining, and other. Thermo scientific pierce high sensitivity streptavidinhrp, 0. For elisa applications, hrp will act on soluble substrates, such as abts or tmb, to yield a colorimetric reaction. Measuring the level of endogenous peptides on ch27 and m12. In contrast to avidin, streptavidin has no carbohydrate and has a mildly acidic pi of 5. Development of a sensitive, rapid, biotinstreptavidin. Thermo scientific streptavidin products use a recombinant form of streptavidin having a mass of 53,000 daltons and a nearneutral pi. Streptavidinap conjugate streptavidinhrp conjugate streptavidinfitc conjugate for immunodetection of biotinylated recombinant proteins catalog nos. Hrp streptavidin conjugate comprises streptavidin as the biotinbinding protein with hrp covalently attached as the enzyme label. Streptavidin hrp conjugates have a very high turnover rate, coupling high sensitivity with short reaction times. Using biotinylated primary or secondary intermediates and peroxidaselabeled streptavidin, antigens can be localized in histological sections, cytospin preparations, or smears. Streptavidin is covalently coupled to alkaline phosphatase ap for the preparation of streptavidin ap conjugate. Streptavidinhrp is useful for detecting biotinylated antibodies, e.
Streptavidin hrp is used with biotinylated proteins and specific chemiluminescent substrates to generate light signal. Streptavidin horseradish peroxidase hrp conjugate catalog no. Thermo scientific pierce high sensitivity streptavidin and neutravidin coated plates enable detection of low concentrations of biotinylated antibodies and other proteins i. Thermo scientific pierce high sensitivity streptavidin hrp conjugate is an exclusive peroxidaseconjugated streptavidin biotinbinding protein that provides signal amplification like polyhrp and exceptional storage stability. Neela parthasarat, eric lecuye, kelly yeeand henry m biotin sp conjugated igg fraction monoclonal mouse anti digoxin 1400 dilution of a 1 m gml stock solution. Specializing in secondary antibodies and conjugates. Prepare a working solution of the streptavidinhrp polymer conjugate by diluting 1. Thermo scientific pierce high sensitivity streptavidinhrp, 2 ml is an exclusive peroxidaseconjugated streptavidin biotinbinding protein that provides signal. This polyhrp conjugate is designed to deliver the highest sensitivity and low background in immunoassays where sample volume is limited or when the target molecule is present at low levels. Just like avidin, streptavidin has very high affinity to biotin molecule, one million times than the common affinity between antigen and antibody. Setadye streptavidinconjugates are extremely bright and photostable and they remain highly fluorescent over a broad ph range ph 4 9. The estimated average number of hrp monomer molecules in sapolyhrp20 conjugate is 100 20 x 5, in sapolyhrp40 200 40 x 5 and in sapolyhrp80 400. Af488streptavidin conjugate streptavidin, alexa fluor.
The detection range 5300 ng was determined using 100. Streptavidin polyhrp is purified to remove unconjugated streptavidin molecules that reduce signal intensity by competing for binding sites with. Specializing in secondary antibodies and conjugates streptavidin. This highly sensitive detector conjugate binds to biotinylated primary or secondary antibodies in enzyme immunoassay, immunoblotting, immunohistochemistry, flow cytometry, hybridoma. Biotinylated antitsh monoclonal antibody mab and hrplabeled streptavidin were first synthesized. Thermo scientific pierce high sensitivity streptavidin hrp conjugate is an exclusive peroxidase conjugated streptavidin biotinbinding protein that provides signal amplification like polyhrp and exceptional storage stability. Streptavidin homotetramers have an extraordinarily high affinity for biotin also known as vitamin b7 or vitamin h. Pierce streptavidin poly hrp is purified to remove unconjugated streptavidin molecules that reduce signal intensity by competing for binding sites with. Streptavidinhrp nel750 and streptavidinap nel751 are highly sensitive detector conjugates. Streptavidin hrp conjugate elisa grade streptavidin has been conjugated to horseradish peroxidase hrp is used with biotinylated antibodies. Streptavidinhrp conjugates have a very high turnover rate, coupling high. Hrp streptavidin conjugate form streptavidin hrp is supplied as a liquid 2 ml aliquot 1.
To assess background from nonspecific binding of conjugate, prepare a similar dilution of the streptavidinhrp polymer conjugate in pbs with 0. Streptavidin protein, hrp conjugate invitrogen 21126. For affinity blotting, blots were probed with hrp pierce or ap millipore conjugated streptavidin for 1 hour at 1. Pierce streptavidin polyhrp is purified to remove unconjugated streptavidin molecules that reduce signal intensity by competing for binding sites with. Streptavidin is a tetrameric bacterial protein isolated from streptomyces avidinii providing 4 high affinity biotin binding sites figure 1. Streptavidinhrp is used with biotinylated proteins and specific chemiluminescent substrates to generate light signal. Ap converts the sensitive substrate xphosphatenbt into a stable, water soluble product. Streptavidin is a 47kd protein that extracted from streptomyces. Thermo scientific pierce high sensitivity streptavidinhrp, prediluted hrp conjugated, prediluted 10gml. A 60kda extracellular protein of streptomyces avidinii with four highaffinity biotin binding sites.
Pierce high sensitivity streptavidin hrp productnumber. Af488streptavidin conjugate, streptavidin, alexa fluor 488 conjugate, s32354 actively helping customers, employees and the global community during the coronavirus sarscov2. The speed and sensitivity of these methods are unequalled by traditional alternatives. The conjugate is provided in liquid form with a proprietary stabilizer added, without preservative. Horseradish peroxidase streptavidin hrp streptavidin is produced by our own coupling procedure that preserves the high specific activity of the peroxidase. Streptavidin polyhrp conjugate is streptavidin biotinbinding protein that is conjugated with polymers of horseradish peroxidase, enabling signal amplification and detection of biotinylated antibodies for ihc and other methods. It does not guarantee that antibody conjugate combinations have been tested for use in the listed applications. Alkaline phosphatase alkaline phosphatase is often used as a label in enzyme immunoassays. Streptavidinap conjugate streptavidinhrp conjugate. Each high sensitivity hrp conjugate is packaged in an easytouse stabilized solution that enables convenient storage at 4c for at least one year. Compared with the avidinbiotinhrp complex abc, hrpconjugated streptavidin is more stable, gives less background, and is more sensitive as reported by shi et al.
The complex and irregular structure of the biotinbinding site makes it highly optimized for biotin binding and confers great speci. Streptavidin has an extremely high binding affinity kd1015 for biotin. Nanocs has developed a unique process to synthesize highly purified streptavdin gold nanoparticles and these products have been distributed through sigma aldrich, fisher scientific and many other companies worldwidely. Jackson immunoresearch laboratories inc catalog no. All tissue sections were treated with dab brown for 10 minutes and counterstained with hematoxylin blue. Pierce high sensitivity streptavidinhrp thermo fisher scientific. Streptavidin binds to biotin and the conjugated hrp provides enzyme activity for detection using an appropriate substrate system. Streptavidin ap conjugate streptavidin hrp conjugate streptavidin fitc conjugate for immunodetection of biotinylated recombinant proteins catalog nos.
Streptavidin poly hrp conjugate is streptavidin biotinbinding protein that is conjugated with polymers of horseradish peroxidase, enabling signal amplification and detection of biotinylated antibodies for ihc and other methods. Incubate the tissue section with the streptavidinperoxidase conjugate for 30 minutes. Streptavidin peroxidase polymer, ultrasensitive s2438. Product data sheet pierce high sensitivity streptavidin. Seta biomedicals streptavidinconjugates are offered with a variety of different seta labels, that span the whole visible and the nir. High sensitivity streptavidin hrp pierce streptavidin. Streptavidinhorseradish peroxidase savhrp is a useful detection enzyme for labeling biotinylatedprimary or seconday antibodies for detection by immunohistochemistry. In situ detection of apoptosis by tunel in fixed frozen, paraffin embedded, or plastic embedded cells and tissues. A gnp conjugated primary antibody and secondary antibody as well as streptavidinhrp streptavidinhorseradish peroxidase were used to improve detection of the analyte.
Unlike avidin, streptavidin has a near neutral isoelectric point and is free of carbohydrate. Dilute streptavidinhrp conjugate according to the manufacturers instructions. Using biotinylated primary or secondary intermediates and alkaline phosphataseconjugated streptavidin, it is suitable for use in solidphase assays, tissuecell staining systems, and blotting applications. In comparison with conventional, high proteinbinding streptavidin horseradish peroxidase savhrp, kpl, 0. Neutravidin protein a specially deglycosylated form of avidin that provides highly specific, lowbackground binding of biotinylated antibodies in many applications. Nanocs streptavidin gold nanoparticle can be used for detection of biotinylated molecules with high sensitivity and specificity. Occasional delays may occur due to unforeseen complexities in the preparation of your product. The goal is to use antibody conjugation with gnp to increase antibody capture, leading to improved analyte binding, induced high sensitive detection. The applications listed are general applications for the conjugate label alone. Alkaline phosphatase streptavidin ap streptavidin is produced by our own coupling procedure that preserves the high specific activity of alkaline phosphatase. Af488streptavidin conjugate streptavidin, alexa fluor 488.
Streptavidinhrp conjugate readytouse reagent for use with the simple western size standard. Thermo scientific pierce streptavidin polyhrp polyhrp. Streptavidinhrp conjugates have a very high turnover rate. Add 100 l of diluted streptavidinhrp per well, cover the microplate and incubate at room temperature for 15 to 45 minutes. A gnp conjugated primary antibody and secondary antibody as well as streptavidin hrp streptavidin horseradish peroxidase were used to improve detection of the analyte. Streptavidinhrp may be diluted in standard diluentassay buffer see table 1 if not otherwise specified by the manufacturer. Thermo scientific pierce high sensitivity neutravidinhrp. Refer to the manufactures data sheet for additional information. Our high sensitivity hrp conjugates outperform the competition on target detection, signal intensity and signaltonoise ratios. Instructions pierce high sensitivity streptavidin coated. For example, experiments indicate that the high sensitivity streptavidin hrp can be diluted eightto 10fold more than traditional hrp conjugates and still produce similar results. Streptavidin hrp conjugate readytouse reagent for use with the simple western size standard.
It is commonly used as a second step reagent for indirect immunofluorescent staining, when used in conjunction with a hrp substrate such as our sensitive fluorogenic adhp peroxidase substrate. Compared with the avidinbiotin hrp complex abc, hrp conjugated streptavidin is more stable, gives less background, and is more sensitive as reported by shi et al. High sensitivity streptavidin and neutravidin coated. Secondary streptavidinhrp conjugate readytouse reagent for jess wes sally sue and peggy sue 2 x 1 ml use with a biotinylated antibody for total protein assay use 042976 and to detect biotinylated molecular weight ladders use 042414 ships with ice pack upon receipt store at 28. Add 100 l of diluted streptavidinhrp per well, cover the microplate and. This polyhrp conjugate is designed to deliver the highest sensitivity and low background in immunoassays where. Hrpstreptavidin conjugate form streptavidinhrp is supplied as a liquid 2 ml aliquot 1. Comparisons of apo and liganded streptavidin crystal structures by weber et al. Thermo scientific pierce high sensitivity streptavidinhrp, pre. Streptavidin poly hrp is purified to remove unconjugated streptavidin molecules that reduce signal intensity by competing for binding sites with.
378 620 509 1114 582 1197 729 1433 613 1653 1513 1561 1271 1433 742 361 98 1229 1209 469 1011 708 1312 173 1585 1081 131 324 1155 1463 1178 1540 1218 1381 504 1577 723 1282 254 602 1174 776 116 627 280 1107 352 1321 1106