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Tnp conjugated bsa

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TNP-conjugated BSA is a protein conjugate consisting of bovine serum albumin (BSA) covalently linked to trinitrophenyl (TNP). It serves as a hapten carrier for immunological studies and applications.

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Lab products found in correlation

Recombinant mouse il 3 and mouse scf, by Thermo Fisher Scientific (2 mentions) Histamine, by Fujifilm (1 mentions) Recombinant mouse il 33, by R&D Systems (1 mentions) Ifnar1, by Jackson ImmunoResearch (1 mentions) Recombinant mouse ifn α, by Miltenyi Biotec (1 mentions) Stat1, by Jackson ImmunoResearch (1 mentions) C57bl 6j mice, by CLEA Japan (1 mentions) Tnp lps, by Biosearch Technologies (1 mentions) Ap conjugated secondary antibody, by Southern Biotech (1 mentions) Isotype specific antibodies, by Southern Biotech (1 mentions) Tnp ficoll, by Biosearch Technologies (1 mentions) Celltrace violet, by Thermo Fisher Scientific (1 mentions) Pnpp substrate, by Merck Group (1 mentions) Ready set go, by Thermo Fisher Scientific (1 mentions) Tnp klh, by Biosearch Technologies (1 mentions) Imject alum, by Thermo Fisher Scientific (1 mentions) Microplate reader, by Molecular Devices (1 mentions)

5 protocols using tnp conjugated bsa

1

Slc15a4-Deficient Mice: A Novel Model

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Slc15a4-deficient mice were generated as previously described (2 (link)). All animal experiments were approved by the Animal Care and Use Committee of the National Center for Global Health and Medicine (NCGM) Research Institute, and were conducted in accordance with institutional procedures. The P815 mouse mastocytoma line was purchased (American Type Culture Collection, VA, USA). The RBL-2H3 rat basophilic leukemia cell line and the anti-TNP mouse IgE (IGELb4) were kind gifts from H. Karasuyama (Tokyo Dental and Medical University). TNP-conjugated BSA (LGC Biosearch Technologies, CA, USA), E. coli LPS and 4-methylumbelliferyl-N-acetyl-β-glucosaminide (4-MUAG) (Sigma, MO, USA), histamine (Wako Pure Chemical Industries, Osaka, Japan), recombinant mouse IL-3 and mouse SCF (Peprotech, NJ, USA) and recombinant mouse IL-33 (R&D Systems, MN, USA) were purchased.
Kobayashi T., Tsutsui H., Shimabukuro-Demoto S., Yoshida-Sugitani R., Karyu H., Furuyama-Tanaka K., Ohshima D., Kato N., Okamura T, & Toyama-Sorimachi N. (2017). Lysosome biogenesis regulated by the amino-acid transporter SLC15A4 is critical for functional integrity of mast cells. International Immunology, 29(12), 551-566.
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2

Deficient Mice for Immune Research

Cited 1 time
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Ifnar1-, Stat1-, and Stat2-deficient mice were obtained from The Jackson Laboratory (Bar Harbor, ME). C57BL/6J mice were purchased from CLEA Japan (Tokyo, Japan). The anti-TNP mouse IgE (IGELb4) was described previously [30 (link)]. TNP-conjugated BSA (LGC Biosearch Technologies, Middlesex, UK, T5050-10) and 4-methylumbellyferyl-N-acetyl-β-glucosaminide (4-MUAG) (Sigma, St. Louis, MO, M2133), recombinant mouse IL-3 and mouse SCF (Peprotech, Rockey Hill, NJ, 213–13 and 250–03, respectively), recombinant mouse Thrombopoietin (Wako Pure Chemicals, Osaka, Japan, 202–19611), recombinant mouse IFN-α (Miltenyi Biotec 130-093-131), and the STAT3 inhibitor S3I-201 (Merck Millipore, Burlington, MA; hereafter MM, 573102) were purchased.
Kobayashi T., Shimabukuro-Demoto S., Tsutsui H, & Toyama-Sorimachi N. (2019). Type I interferon limits mast cell–mediated anaphylaxis by controlling secretory granule homeostasis. PLoS Biology, 17(11), e3000530.
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3

Mouse Immunization and Antibody Response Assay

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4–6-wk-old mice were immunized i.p. with 50 µg OVA with Alum (Sigma-Aldrich) in PBS at day 0, boosted at day 14, and bled at days 0 and 21. Mice were immunized i.p. with 25 µg TNP-LPS (Biosearch Technologies) in PBS or with 25 µg TNP–Ficoll (Biosearch Technologies) at day 0 and bled at days 0 and 14. Antigen-specific antibody responses were analyzed by OVA- or TNP-specific ELISA using 96-well plates coated with either OVA- or TNP-conjugated BSA (Biosearch Technologies) at 10 µg/ml in PBS. For serum Ig levels, plates were coated with isotype-specific antibodies (SouthernBiotech). AP-conjugated secondary antibodies (SouthernBiotech) and PNPP (Sigma-Aldrich) were used for ELISA.
Mooster J.L., Le Bras S., Massaad M.J., Jabara H., Yoon J., Galand C., Heesters B.A., Burton O.T., Mattoo H., Manis J, & Geha R.S. (2015). Defective lymphoid organogenesis underlies the immune deficiency caused by a heterozygous S32I mutation in IκBα. The Journal of Experimental Medicine, 212(2), 185-202.
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4

Measuring Antibody Responses in Mice

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Basal serum Ig titers were quantified by ELISA by using HRP-conjugated Ig from Southern Biotech (Victoria, Australia). To evaluate T cell-dependent or T cell-independent antibody responses, mice were administered intraperitoneal doses of the following: 100 μg of TNP-KLH in alum; 50 μg of TNP-Ficoll; or 50 μg of TNP-LPS. Titers of antibodies to TNP were measured by ELISA with plate-bound TNP-conjugated BSA (Biosearch Technologies) and isotype-specific horseradish peroxidase-conjugated secondary antibodies (Southern Biotech).
Hanihara-Tatsuzawa F., Miura H., Kobayashi S., Isagawa T., Okuma A., Manabe I, & MaruYama T. (2014). Control of Toll-like Receptor-mediated T Cell-independent Type 1 Antibody Responses by the Inducible Nuclear Protein IκB-ζ. The Journal of Biological Chemistry, 289(45), 30925-30936.
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5

Antigen-Specific Antibody Response in Mice

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Two-month-old mice were immunized intraperitoneally with 10 μg of TNP-KLH (Biosearch Technologies, Petaluma, Calif) and Imject Alum (Thermo Fisher Scientific). Mice were subsequently boosted on day 14 with 2.5 μg of TNP-KLH and Imject Alum. Splenocytes were harvested on day 21, labeled with CellTrace Violet (Life Technologies) and cultured at a concentration of 3 × 105 cells in the presence of media alone or KLH at the indicated concentration. On day 3, supernatants were harvested, and IL-2 and IFN-γ levels were measured by means of ELISA (Ready-SET-Go!; eBioscience). Levels of antigen-specific antibody responses from serum obtained on days 0 and 21 were analyzed by using TNP-specific ELISA with 96-well plates coated with TNP-conjugated BSA (Biosearch Technologies) at 10 μg/mL in PBS. Plates were coated with TNP/BSA conjugated at a molar ratio of 2:1 to detect high-affinity antibody and TNP/BSA at a molar ratio of 32:1 to detect low-affinity antibody. Sera were diluted at 1:300, 1:1,000, 1:3,000, 1:9,000, 1:27,000, and 1:81,000. KLH-specific antibodies were detected with IgG–alkaline phosphatase. PNPP Substrate (Sigma) was added, and OD 405 was determined with a microplate reader (Molecular Devices).
Platt C.D., Chou J., Houlihan P., Badran Y.R., Kumar L., Bainter W., Poliani P.L., Perez C.J., Dent S.Y., Clapham D.E., Benavides F, & Geha R.S. (2017). Leucine-rich repeat containing 8A (LRRC8A)–dependent volume-regulated anion channel activity is dispensable for T-cell development and function. The Journal of allergy and clinical immunology, 140(6), 1651-1659.e1.
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