Products
- Amino-Dextrans (Amdex)
- Mono Amino Dextrans (mAmDex)
- Amino-Ficoll
- TNP-Ficoll
- TNP20-BSA
- Anti-Ig Dextran Conjugates
- Peroxidase Immunoreagents
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AmDex (Amino dextran, AECM dextran) is prepared from dextran using a protocol yielding a stable, amino functionalized dextran. The molecular weight profile is essentially unchanged, as determined by analytical size exclusion chromatography (Figure 1). We do extensive processing and testing to ensure that free amino reagent is removed. This approach results in a consistent product with minimal lot-to-lot variation.
Each batch of amino-dextran polymer is evaluated for molecular weight distribution and for the amine content, expressed as amines/polymer. A certificate of analysis is provided for each lot.
Custom polymer preparations are available.
Figure 1 Size exclusion chromatography of 70 kDa dextrans
Overlay of dextran and AmDex
Products: 70 kDa Dextran
100 mg $95.00
200 mg $175.00
500 mg $350.00
Bulk On request
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mAmDex (monoamino dextran, mAECM dextran) is prepared by proprietary methods yielding a stable, monoamino functionalized dextran . The molecular weight profile is essentially unchanged, as determined by analytical size exclusion chromatography . Extensive processing and testing is performed to ensure that free amino reagent is removed. This approach results in a consistent product with minimal lot-to-lot variation.
mAmDex is unique in that it has a single amino group at one end of the polymer. Much like PEG, it can be used to increase protein half-life, reduce immunogenicity, and passivate surfaces.
Mono-carboxyl dextran and custom molecular weight polymer preparations are available upon request.
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| Figure 1 Mono-amino dextran | Figure 2 Mono carboxyl dextran |
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As a T-cell independent carrier (TI carrier), coupling Ficoll with haptens has been shown to produce robust antigen-specific primary immune memory (Vos, Q., Lees, A., Wu, Z. Q., Snapper, C. M., and Mond, J. J. (2000) B-Cell Activation by T-Cell-Independent Type 2 Antigens as an Integral Part of the Humoral Immune Response to Pathogenic Microorganisms. Immunol. Rev. 176, 154-170.) Our method for amino functionalization yields stable primary amine groups. We carefully control the amine:polymer ratio and process to remove excess amine reagent. Custom Ficoll preparations are available upon request. GE Healthcare’s trademark Ficoll is a highly branched, 400kDa, hydrophilic polysaccharide widely used for blood separation.
Carboxyl functionalized Ficoll and custom orders available upon request.
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When conjugated to Ficoll, the TNP-Ficoll complex can be used as a T cell independent antigen in the study of B cell lymphocyte activation. 2,4,6-Trinitrophenyl (TNP) carries an absorbance between 330-430nm with peaks at 348 and 415nm. Molar extinction coefficients range between 10,000-15,000 M-1 cm-1.
Custom sizes, ratios, and molecular weights are available upon request.
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| Figure 1 Chemical structure of TNP Conjugated to polysaccharide | Figure 2 Wavelength scan of 0.2mg/mL TNP-Ficoll400×180. Peak points at 348nm and 415nm. |
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TNP-BSA can be used in tandem with TNP-Ficoll for immunoassays with anti-TNP antisera among other applications. (Arnold, C. et al., Mutagenetix http://mutagenetix.utsouthwestern.edu:80, 2010). 2,4,6-Trinitrophenyl (TNP) carries an absorbance between 330-430nm with peaks at 348 and 415nm. Molar extinction coefficients range between 10,000-15,000 M?1 cm?1.
Bovine Serum Albumin (BSA) Fraction V is a 66.5kDa protein commonly used in immunochemical applications such as ELISA IHC, and immunoblots. It is widely used as a quantification standard for proteins and as an enzyme stabilizer during DNA digestion. BSA has an extinction coefficient of 44,000 M-1 cm-1 at 280nm.
Custom sizes, ratios, and molecular weights are available upon request.
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| Figure 1 Chemical structure of TNP Conjugated to BSA | Figure 2 Wavelength scan of TNP20-BSA Peak points at 348nm and 415nm |
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FinaBio#0001 Anti-IgD dextran (mouse) 10 ug/vial
FinaBio#0002 Anti-IgD dextran (human) 10 ug/via
FinaBio#0003 Anti-IgM dextran (mouse) 10 ug/vial
FinaBio#0004 Anti-igM dextran (human) 10 ug/vial
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Crosslinking membrane IgD on B cells with unconjugated anti-IgD antibody induces B cell proliferation at ug/ml concentrations. By contrast, dextran-anti-IgD antibodies (alpha-delta-dex) induce vigorous B cell proliferation at ng/ml concentrations (Brunswick et al., J. Immunol. 140:3364, 1988). Further, in contrast to unconjugated anti-IgD, alpha-delta-dex-activated B cells undergo substantial Ig class switching and differentiation into Ig-secreting cells in the presence of various second signals, including cytokines and Toll-like receptor ligands (Snapper and Mond, J. Immunol. 157:2229, 1996). Thus, alpha-delta-dex is an efficient and potent polyclonal B cell activator for studying a wide range of B cell functions.
Figure 1 Uptake of 3H thymidine as a marker of B cell activation
References
J. Immunol. 140:3364, 1988. M. Brunswick et al.
Picogram quantities of anti-Ig antibodies coupled to dextran induce B cell proliferation. To investigate the properties which enable type 2 Ag, as exemplified by dextran and Ficoll, to stimulate high levels of antibody responses in the relative absence of T cells, we conjugated anti-IgD and anti-IgM mAb to both dextran and Ficoll and examined their B cell-activating properties. Such conjugated anti-Ig antibodies stimulated both early and later stages of B cell activation at picogram concentrations, which are at least 1000-fold lower than that required for B cell stimulation by unconjugated anti-Ig antibodies, and the level of proliferation they stimulated was on average 10-fold greater. Furthermore, concentrations of anti-Ig dextran (100 pg/ml) which modulated little sIgD from the B cell surface were strong inducers of enhanced B cell expression of MHC class II molecules. Conjugation of Fab fragments of anti-IgD or nonmitogenic anti-IgM mAb to dextran rendered them as mitogenic as dextran conjugated to strongly stimulatory anti-IgD or anti-IgM antibodies. The ability of dextran and Ficoll to serve as effective carrier molecules for anti-IgD was not related solely to their large m.w., because anti-IgD coupled to polymerized BSA (m.w. 1.5 X 10(6), was only 10- to 50-fold more potent than unconjugated anti-IgD antibodies at stimulating B cell DNA synthesis. These results suggest, therefore, that the unique ability of picogram concentrations of haptenated type 2 Ag to stimulate Ig secretion in the absence of T cells may be a function of their ability to promote effective cross-linking without resulting in the modulation of sIg. This would enable such Ag to mediate repetitive B cell signaling, a situation that cannot be achieved by unconjugated anti-Ig antibodies which result in modulation of sIg at their mitogenic concentrations. These compounds therefore may be employed to study B cell activation stimulated by sIg cross-linking at concentrations that may more closely reflect those which are achieved under physiologic conditions by type 2 Ag.
J. Immunol. 157: 2229, 1996. C. M. Snapper and J. J. Mond.
A model for induction of T cell-independent humoral immunity in response to polysaccharide antigens. Abstract. We provide a model for induction of T cell-independent, polysaccharide-specific Ig secretion in response to bacterial challenge. Two predominant pathways are defined that require the concerted action of multivalent membrane Ig cross-linking by the polysaccharide Ag with 1) various B cell-activating moieties contained within the bacterial pathogen and/or 2) cytokines, such as IFN-gamma and granulocyte-macrophage colony-stimulating factor produced by NK cells and macrophages, that become activated in a T cell-independent manner during bacterial infection.
Macrophage colony-stimulating factor, show synergistic stimulatory activity in inducing Ig secretion in B cells stimulated by a multivalent ligand that mimics TI-2 antigens. The recent finding that natural killer cells have receptors for various classes of polysaccharides supports a role for these cells in regulating responses to TI-2 antigens.
PolyHRP Immunoassay Conjugates
Antibodies are affinity purified from hyperimmune goat serum. Horseradish peroxidase is polymerized through proprietary techniques and then covalently linked to the immunoglobulin for use in immunoassays or histo/cytochemical procedures.
Pricing for all PNs:
100µL: $65
500µL: $150
1mL: $220
Bulk on Request
Custom conjugation available upon request.
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SBP-XP™ Immunoassay Conjugates
Soybean peroxidase is relatively new to the immunoreagent market but can be used interchangeably with HRP substrates. SBP is a single isozyme with a more tightly bound heme group than HRP, making it more thermostable. The enzyme retains its activity to temperatures up to 75°C, over a wider pH range, as well as less lot to lot variability. In chemiluminescence the SBP produces a “glow” rather than the ‘flash” seen with HRP and can therefore be read up to 2 hours or more after substrate addition.
Antibodies are affinity purified from hyperimmune goat serum. Soybean peroxidase is covalently linked to the immunoglobulin or streptavidin for use in immunoassays or histo/cytochemical procedures.
Pricing for all PNs:
100µL: $65
500µL: $150
1mL: $220
Bulk on Request
Custom conjugation available upon request.
| Figure 1 A clear plate was coated with mouse IgG at 100ng/mL. Biotinylated Gt-a-Mouse IgG was serially diluted 1:3 from 200ng/mL and incubated for 30 min. The conjugates were used at 20ng/mL and incubated for 30 minutes before addition TMB substrate.
Aliquots of SBP-XP™ Streptavidin were held at 4°C, RT, and 37°C for three months. |
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SBP-XP is a trademark of IBEX Technologies Inc.
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