Antibodies against mouse CD4, CD19, CD95, GL-7, ICOS, ICOSL, PD-1, and B220-647 were purchased from Biolegend, CXCR5, B220, IgG1, IgG2a, IgG2b, and IgG3 antibodies from BD Biosciences, Bcl-6, XBP-1, and IRF-4 from Santa Cruz, and BAFF (1C9) from Enzo. Anti-μ F(ab)2 was purchased from Jackson ImmunoResearch. Anti-μ (clone B7.6), anti-NP (clones H33L and B1-8), 2.4G2 (FcγRIIb /FcγRIII block) and Ac38 idiotype antibodies were purified from hybridoma supernatants. (Ac38 is an idiotype antibody that recognizes B-1-8 specificities generated during NP immunization). Recombinant mouse BR3-Fc and isotype control protein were generated using mammalian expression systems and standard purification protocols. H33L and B1-8 were gifts from Dr. Garnett Kelsoe (Duke University). IL-4 and IL-5 were purchased from Peprotech, recombinant BAFF (rBAFF) from R&D Systems, NP-OSu from Biosearch Technologies, KLH and PNA-biotin from Sigma Aldrich, alum from Thermo Scientific, and streptavidin-Alexa 488 and Alexa 647 from Invitrogen. Streptavidin-alkaline phosphatase and anti-IgG alkaline phosphatase were purchased from Southern Biotech. The Fc-binding TG19320 peptide was synthesized as described (39 (link), 40 (link)).
Anti igg alkaline phosphatase
Manufactured by Southern Biotech
Anti-IgG alkaline phosphatase is a laboratory reagent used to detect and quantify the presence of immunoglobulin G (IgG) in a sample. It functions as a conjugate, binding to IgG and producing a measurable signal through the enzymatic activity of alkaline phosphatase.
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Lab products found in correlation
Streptavidin alexa 488, by Thermo Fisher Scientific (1 mentions)
Icosl, by BioLegend (1 mentions)
Streptavidin alkaline phosphatase, by Southern Biotech (1 mentions)
Np osu, by Biosearch Technologies (1 mentions)
Vector blue, by Vector Laboratories (1 mentions)
Multiscreen hts plate, by Merck Group (1 mentions)
Immunospot analyzer, by Cellular Technology (1 mentions)
Roswell park memorial institute (rpmi), by Thermo Fisher Scientific (1 mentions)
2 protocols using anti igg alkaline phosphatase
1
Antibody Reagents for B Cell Immunology
2
Influenza Virus-Specific Antibody ELISA
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MultiScreen HTS plates (Millipore) were coated with 50 µl/well of recombinant HA protein at a concentration of 2 µg/ml in PBS overnight at 4°C. The coating solution was removed, and plates were washed six times with PBS-T followed by incubation with 200 µl of complete medium (10% fetal bovine serum and 1% L-glutamine, penicillin, and streptomycin in RPMI [Life Technologies]) for 2 h at 37°C. Subsequently, bone marrow cells from mice infected with the PR8 influenza virus were isolated and plated (3 × 106 cells/well) in duplicate in complete medium for 4–5 h at 37°C. Plates were then washed six times with PBS-T followed by addition of secondary antibody (50 µl) in blocking solution (0.5% BSA in PBS; anti-IgG–alkaline phosphatase; Southern Biotech [1030–04]) to each well, then incubated overnight at 4°C. The plates were washed three times with PBS-T and three times with PBS. Spots were developed with Vector Blue (Vector Laboratories) and quantified using an ImmunoSpot analyzer (Cellular Technology Limited).
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