Alkaline phosphatase conjugated goat anti mouse total ig secondary ab

Manufactured by Southern Biotech

Alkaline Phosphatase-conjugated goat anti-mouse total Ig secondary Ab is a laboratory reagent used for the detection of mouse immunoglobulins (Ig) in various immunoassay applications. It contains alkaline phosphatase enzyme-labeled antibodies that bind to the constant regions of mouse Ig molecules, allowing for colorimetric or chemiluminescent detection.

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Lab products found in correlation

Multiscreen ha mixed cellulose ester membrane plates, by Merck Group (5 mentions) Bcip nbt substrate, by Merck Group (3 mentions) Nbt bcip substrate nitroblue tetrazolium 5 bromo 4 chloro 3 indolyl phosphate, by Merck Group (2 mentions) 5 bromo 4 chloro 3 indolyl phosphate nitro blue tetrazolium, by Merck Group (1 mentions)

Close spelling variants of this product

Goat anti-mouse total Ig Goat anti-mouse Ig Alkaline phosphatase

5 protocols using alkaline phosphatase conjugated goat anti mouse total ig secondary ab

Quantifying Anti-GPI Antibody Secreting Cells

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Anti-GPI antibody secreting cells were measured by ELISpot as described (25 (link)). Briefly, cells from the joint draining lymph nodes (axillary, brachial, and popliteal LNs) from 6 week-old KRN.g7, IDO2 ko KRN.g7, R235W KRN.g7, and Y346X KRN.g7 mice were plated at 4 × 10⁵ cells per well and diluted serially 1:4 in Multiscreen HA mixed cellulose ester membrane plates (Millipore) coated with histidine (his)-tagged glucose-6-phosphate isomerase (GPI) (10μg/ml). The cells were incubated on the GPI-coated plates for 4hr at 37°C. The Ig secreted by the plated cells was detected by Alkaline Phosphatase-conjugated goat anti-mouse total Ig secondary Ab (Southern Biotechnology Associates) and visualized using NBT/BCIP substrate (nitroblue tetrazolium / 5-bromo-4-chloro-3-indolyl phosphate; Sigma).

Merlo L.M., Peng W., DuHadaway J.B., Montgomery J.D., Prendergast G.C., Muller A.J, & Mandik-Nayak L. (2021). The Immunomodulatory Enzyme IDO2 Mediates Autoimmune Arthritis Through a Non-Enzymatic Mechanism. Journal of immunology (Baltimore, Md. : 1950), 208(3), 571-581.

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Quantifying Anti-GPI Antibody Secreting Cells by ELISpot

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Anti-GPI antibody secreting cells were measured by ELISpot as described (36 (link)). Briefly, cells from the joint draining lymph nodes (axillary, brachial, and popliteal LNs) from 6 week-old KRN.g7, IDO1 ko KRN.g7, IDO2 ko KRN.g7, and dko KRN.g7 mice were plated at 4 × 10⁵ cells per well and diluted serially 1:4 in Multiscreen HA mixed cellulose ester membrane plates (Millipore) coated with GPI-his (10 μg/ml). The cells were incubated on the Ag-coated plates for 4 h at 37°C. The Ig secreted by the plated cells was detected by Alkaline Phosphatase-conjugated goat anti-mouse total Ig secondary Ab (Southern Biotechnology Associates) and visualized using NBT/BCIP substrate (nitroblue tetrazolium/5-bromo-4-chloro-3-indolyl phosphate; Sigma).

Merlo L.M., DuHadaway J.B., Montgomery J.D., Peng W.D., Murray P.J., Prendergast G.C., Caton A.J., Muller A.J, & Mandik-Nayak L. (2020). Differential Roles of IDO1 and IDO2 in T and B Cell Inflammatory Immune Responses. Frontiers in Immunology, 11, 1861.

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ELISPOT Assay for Ig-Secreting Cells

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Cells from the joint draining LN (axillary, brachial, and popliteal LNs) from 6 week-old control mouse Ig or IDO2 Ig-treated KRN.g7 mice were plated at 4 × 10⁵ cells per well and diluted serially 1:4 in Multiscreen HA mixed cellulose ester membrane plates (Millipore) coated with GPI-his (10μg/ml). The cells were incubated on the Ag-coated plates for 4hr at 37ºC. The Ig secreted by the plated cells was detected by Alkaline Phosphatase-conjugated goat anti-mouse total Ig secondary Ab (Southern Biotechnology Associates) and visualized using NBT/BCIP substrate (nitroblue tetrazolium / 5-bromo-4-chloro-3-indolyl phosphate; Sigma).

Merlo L.M., Grabler S., DuHadaway J.B., Pigott E., Manley K., Prendergast G.C., Laury-Kleintop L.D, & Mandik-Nayak L. (2017). Therapeutic Antibody Targeting of Indoleamine-2,3-Dioxygenase (IDO2) Inhibits Autoimmune Arthritis. Clinical immunology (Orlando, Fla.), 179, 8-16.

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Quantifying Antigen-specific B Cells

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Cells from the joint draining LN (axillary, brachial, and popliteal LNs) from 6 week-old KRN.g7, IDO1 ko KRN.g7, and IDO2 ko KRN.g7 mice were plated at 4 × 10⁵ cells per well and diluted serially 1:4 in Multiscreen HA mixed cellulose ester membrane plates (Millipore) coated with GPI-his (10μg/ml). The cells were incubated on the Ag-coated plates for 4hr at 37°C. The Ig secreted by the plated cells was detected by Alkaline Phosphatase-conjugated goat anti-mouse total Ig secondary Ab (Southern Biotechnology Associates) and visualized using NBT/BCIP substrate (nitroblue tetrazolium / 5-bromo-4-chloro-3-indolyl phosphate; Sigma).

Merlo L.M., Pigott E., DuHadaway J.B., Grabler S., Metz R., Prendergast G.C, & Mandik-Nayak L. (2014). IDO2 is a critical mediator of autoantibody production and inflammatory pathogenesis in a mouse model of autoimmune arthritis. Journal of immunology (Baltimore, Md. : 1950), 192(5), 2082-2090.

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Quantifying Mouse Antibody-Secreting Cells

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LN cells were plated at 4 × 10⁵ cells per well and diluted serially 1:4 in Multiscreen HA mixed cellulose ester membrane plates (Millipore) coated with GPI-his (5µg/ml). The cells were incubated on the Ag-coated plates for 4h at 37°C. The Ig secreted by the plated cells was detected by Alkaline Phosphatase-conjugated goat anti-mouse total Ig secondary Ab (Southern Biotechnology Associates) and visualized using NBT/BCIP substrate (nitroblue tetrazolium / 5-bromo-4-chloro-3-indolyl phosphate; Sigma).

Pigott E., DuHadaway J.B., Muller A.J., Gilmour S., Prendergast G.C, & Mandik-Nayak L. (2014). 1-Methyl-Tryptophan Synergizes with Methotrexate to Alleviate Arthritis in a Mouse Model of Arthritis. Autoimmunity, 47(6), 409-418.

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