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Streptavidin sa alkaline phosphatase

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Streptavidin (SA)-alkaline phosphatase is a conjugate that combines the high-affinity binding of streptavidin to biotin with the enzymatic activity of alkaline phosphatase. It is used as a detection reagent in various biotechnological applications.

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Lab products found in correlation

Alkaline phosphatase conjugated anti mouse igg, by Thermo Fisher Scientific (8 mentions) Biotinylated anti kappa ab, by Thermo Fisher Scientific (8 mentions) Vector blue alkaline phosphatase substrate kit 3, by Vector Laboratories (6 mentions) Biotinylated anti mouse igm, by Jackson ImmunoResearch (6 mentions) Multiscreen 96 well filtration plate, by Merck Group (2 mentions) Anti igm or anti igg, by Thermo Fisher Scientific (2 mentions) Computerized imaging analysis system, by Cellular Technology (2 mentions) Nucleosome, by Merck Group (2 mentions) Salmon sperm dsdna, by Thermo Fisher Scientific (2 mentions) Nucleosome, by Thermo Fisher Scientific (2 mentions) Smrnp, by Thermo Fisher Scientific (2 mentions) Elispot plate imaging analysis system, by Cellular Technology (2 mentions) Calf thymus histone, by Merck Group (2 mentions) Nucleosome coated plates, by Merck Group (2 mentions) Sa alkaline phosphatase or alkaline phosphatase conjugated anti mouse igg, by Thermo Fisher Scientific (2 mentions) Elispot plate imaging and analysis system, by Cellular Technology (2 mentions) Biotinylated igg1, by Southern Biotech (2 mentions) Biotinylated igg2b, by Southern Biotech (2 mentions) Anti igm or anti igg capture antibodies, by Thermo Fisher Scientific (2 mentions) Ap igg2c abs, by Southern Biotech (2 mentions)

Spelling variants of this product

Streptavidin-alkaline phosphatase (24 citations) Alkaline Phosphatase (4 citations) SA-alkaline phosphatase (2 citations) Streptavidin-alkaline phosphatase (Vector SA-5100 (2 citations)

10 protocols using streptavidin sa alkaline phosphatase

1

Quantification of Antibody-Secreting Cells by ELISpot

Cited 1 time
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ELISpot assays were performed as previously described (16 (link)). Briefly, splenocytes in 10% RPMI were plated at a concentration of 1 × 105 cells/well onto anti-IgM or anti-IgG (Invitrogen, Grand Island, NY) coated, or at 1 × 106 cells/well on dsDNA-, histone-, or nucleosome-coated multiscreen 96-well filtration plates (Millipore, Bedford, MA). Serially diluted (1:2) cells were incubated for 6 h at 37°C. IgM-producing AFCs were detected using biotinylated anti-mouse IgM (Jackson Immunoresearch, West Grove, PA) and streptavidin (SA)-alkaline phosphatase (Vector Laboratories, Burlingame, CA). IgG-producing AFCs were detected using alkaline phosphatase-conjugated anti-mouse IgG (Molecular Probes, Grand Island, NY). dsDNA-, histone-, and nucleosome-specific AFCs were detected by biotinylated anti-kappa Ab (Invitrogen, Grand Island, NY) and streptavidin (SA)-alkaline phosphatase (Vector Laboratories, Burlingame, CA) or alkaline phosphatase-conjugated anti-mouse IgG (Molecular Probes, Grand Island, NY). Plates were developed using the Vector Blue Alkaline phosphatase Substrate Kit III (Vector Laboratories, Burlingame, CA). ELISpots were enumerated and analyzed using a computerized imaging/analysis system (Cellular Technology, Shaker Heights, OH).
Wong E.B., Soni C., Chan A.Y., Domeier P.P., S.h.w.e.t.a.n.k., Abraham T., Limaye N., Khan T.N., Elias M.J., Chodisetti S.B., Wakeland E.K, & Rahman Z.S. (2015). B cell-intrinsic CD84 and Ly108 maintain germinal center B cell tolerance. Journal of immunology (Baltimore, Md. : 1950), 194(9), 4130-4143.
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2

Quantifying Antibody-Producing Cells via ELISpot

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ELISpot assays were performed as described (37 (link)). Briefly, splenocytes in 10% RPMI with antibiotics were plated at a concentration of 1 × 105 cells/well onto anti-IgM or anti-IgG (Invitrogen, Grand Island, NY) coated, or at 1 × 106 cells/ well on dsDNA-, histone-, or nucleosome-coated multiscreen 96-well filtration plates (Millipore, Bedford, MA). Serially diluted (1:2) cells were incubated for 6 h at 37°C. IgM-producing AFCs were detected using biotinylated anti-mouse IgM (Jackson Immunoresearch, West Grove, PA) and streptavidin (SA)-alkaline phosphatase (Vector Laboratories, Burlingame, CA). IgG-producing AFCs were detected using alkaline-phosphatase conjugated anti-mouse IgG (Molecular Probes, Grand Island, NY). dsDNA-, histone-, and nucleosome-specific AFCs were detected by biotinylated anti-kappa Ab (Invitrogen, Grand Island, NY) and streptavidin (SA)-alkaline phosphatase (Vector Laboratories, Burlingame, CA). Plates were developed using the Vector Blue Alkaline-phosphatase Substrate Kit III (Vector Laboratories, Burlingame, CA). ELISpots were counted and analyzed using a computerized imaging/ analysis system (Cellular Technology, Shaker Heights, OH).
Soni C., Wong E.B., Domeier P.P., Khan T.N., Satoh T., Akira S, & Rahman Z.S. (2014). B cell-intrinsic TLR7 signaling is essential for the development of spontaneous germinal centers. Journal of immunology (Baltimore, Md. : 1950), 193(9), 4400-4414.
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3

Quantitative ELISpot Assay for Autoantibodies

Cited 2 times
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ELISpot assays were performed as previously described (5 (link), 6 (link)). Briefly, splenocytes in RPMI containing 10% fetal bovine serum were plated at a concentration of 1 x 106 cells/well onto salmon sperm dsDNA- (Invitrogen), nucleosome- (histone from Sigma Aldrich plated on a layer of dsDNA coating)- or SmRNP- (Arotec Diagnostics) coated multiscreen 96-well filtration plates (Millipore, Bedford, MA). Serially diluted (1:2) cells were incubated for 12 h at 37°C. dsDNA-, nucleosome-, and SmRNP-specific AFCs were detected by biotinylated anti-kappa Ab (Invitrogen) and streptavidin (SA)-alkaline phosphatase (Vector Laboratories) or alkaline phosphatase-conjugated anti-mouse IgG (Molecular Probes). Plates were developed using the Vector Blue Alkaline phosphatase Substrate Kit III (Vector Laboratories). ELISpots were enumerated and analyzed using a computerized ELISpot plate imaging/ analysis system (Cellular Technology).
Chodisetti S.B., Fike A.J., Domeier P.P., Schell S.L., Mockus T.E., Choi N.M., Corradetti C., Hou B., Atkins H.M., Caricchio R., Decker T., Lukacher A.E., Olsen N, & Rahman Z.S. (2020). Serine phosphorylation of the STAT1 transactivation domain promotes autoreactive B cell and systemic autoimmunity development. Journal of immunology (Baltimore, Md. : 1950), 204(10), 2641-2650.
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4

ELISpot Assay for Detecting Autoantibodies

Cited 2 times
Check if the same lab product or an alternative is used in the 5 most similar protocols
ELISpot assays were performed as previously described (25 (link), 43 (link)). Briefly, splenocytes in RPMI containing 10% fetal bovine serum were plated at a concentration of 1 × 106 cells/well onto salmon sperm dsDNA- (Invitrogen), nucleosome- (histone from Sigma Aldrich plated on a layer of dsDNA coating)- or SmRNP- (Arotec Diagnostics) coated multiscreen 96-well filtration plates (Millipore, Bedford, MA). Serially diluted (1:2) cells were incubated for 12 h at 37°C. dsDNA-, nucleosome-, and SmRNP-specific AFCs were detected by biotinylated anti-kappa Ab (Invitrogen) and streptavidin (SA)-alkaline phosphatase (Vector Laboratories) or alkaline phosphatase-conjugated anti-mouse IgG (Molecular Probes). Plates were developed using the Vector Blue Alkaline phosphatase Substrate Kit III (Vector Laboratories). ELISpots were enumerated and analyzed using a computerized ELISpot plate imaging/ analysis system (Cellular Technology).
Chodisetti S.B., Fike A.J., Domeier P.P., Singh H., Choi N.M., Corradetti C., Kawasawa Y.I., Cooper T.K., Caricchio R, & Rahman Z.S. (2020). Type II but not type I interferon signaling is indispensable for TLR7-promoted development of autoreactive B cells and systemic autoimmunity. Journal of immunology (Baltimore, Md. : 1950), 204(4), 796-809.
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5

ELISpot Assay for Antibody-Producing Cells

Cited 1 time
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ELISpot assays were performed as described (Wong et al., 2012 (link)). Briefly, splenocytes in RPMI containing 10% fetal bovine serum were plated at a concentration of 1 × 105 cells/well onto anti-IgM-, anti-IgG-, salmon sperm dsDNA- (Invitrogen, Grand Island, NY), calf thymus histone- (Sigma Aldrich, St. Louis, MO), or nucleosome-coated plates (Millipore, Bedford, MA). Serially diluted (1:2) cells were incubated for 6 hr at 37°C. IgM-producing AFCs were detected using biotinylated anti-mouse IgM (Jackson ImmunoResearch, West Grove, PA) and streptavidin (SA)-alkaline phosphatase (Vector Laboratories, Burlingame, CA). IgG-producing AFCs were detected using alkaline phosphatase-conjugated anti-mouse IgG (Molecular Probes, Grand Island, NY). dsDNA-, histone-, and nucleosome-specific AFCs were detected by biotinylated anti-kappa Ab (Invitrogen, Grand Island, NY) and SA-alkaline phosphatase or alkaline phosphatase-conjugated anti-mouse IgG (Molecular Probes, Grand Island, NY). Plates were developed using the Vector Blue Alkaline phosphatase Substrate Kit III (Vector Laboratories, Burlingame, CA). ELISpots were enumerated using a computerized ELISpot plate imaging and analysis system (Cellular Technology, Shaker Heights, OH).
Domeier P.P., Chodisetti S.B., Schell S.L., Kawasawa Y.I., Fasnacht M.J., Soni C, & Rahman Z.S. (2018). B-Cell-Intrinsic Type 1 Interferon Signaling Is Crucial for Loss of Tolerance and the Development of Autoreactive B Cells. Cell reports, 24(2), 406-418.
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6

Quantifying Mouse Autoantibody Isotypes by ELISA

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ELISA plates were coated with anti-IgM or anti-IgG capture antibodies (from Invitrogen, Grand Island, NY) and detected using biotinylated anti-mouse IgM (Jackson Immunoresearch, West Grove, PA) or alkaline-phosphatase conjugated anti-mouse IgG (Molecular Probes, Grand Island, NY). Total IgG autoAb titers were measured in ELISA plates coated with dsDNA, histone, nucleosome, Sm/RNP or cardiolipin and detected with biotinylated anti-kappa Ab (Invitrogen, Grand Island, NY). IgG subtype-specific autoAb titers were detected by biotinylated-IgG1, biotinylated IgG2b, and AP-IgG2c Abs (Southern Biotech, Birmingham, AL). Biotinylated antibodies were detected by streptavidin (SA)-alkaline phosphatase (Vector Laboratories, Burlingame, CA). The plates were developed by the PNPP (p-Nitrophenyl Phosphate, Disodium Salt) (Thermo Fisher Scientific, Rockford, IL) substrates for alkaline phosphatase.
Wong E.B., Soni C., Chan A.Y., Domeier P.P., S.h.w.e.t.a.n.k., Abraham T., Limaye N., Khan T.N., Elias M.J., Chodisetti S.B., Wakeland E.K, & Rahman Z.S. (2015). B cell-intrinsic CD84 and Ly108 maintain germinal center B cell tolerance. Journal of immunology (Baltimore, Md. : 1950), 194(9), 4130-4143.
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7

Serum Autoantibody ELISA Assay

Cited 1 time
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Serum autoAbs were measured using standard ELISA protocols as described (3 (link)). Briefly, total IgG autoAb titers were measured in ELISA plates coated with salmon sperm dsDNA, nucleosome or smRNP and detected with biotinylated secondary Ab followed by streptavidin (SA)-alkaline phosphatase (Vector Laboratories). The plates were developed using PNPP (p-Nitrophenyl Phosphate, Disodium Salt) (Thermo Fisher Scientific) substrates for alkaline phosphatase and read at λ405 nm on Synergy H1 (BioTek Instruments).
Chodisetti S.B., Fike A.J., Domeier P.P., Singh H., Choi N.M., Corradetti C., Kawasawa Y.I., Cooper T.K., Caricchio R, & Rahman Z.S. (2020). Type II but not type I interferon signaling is indispensable for TLR7-promoted development of autoreactive B cells and systemic autoimmunity. Journal of immunology (Baltimore, Md. : 1950), 204(4), 796-809.
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8

Serum Autoantibody Detection by ELISA

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Serum autoAbs were measured using standard ELISA protocols as described (16 (link)). Briefly, total IgG autoAb titers were measured in ELISA plates coated with salmon sperm dsDNA, nucleosome or smRNP and detected with biotinylated secondary Ab followed by streptavidin (SA)-alkaline phosphatase (Vector Laboratories). Plates were developed using PNPP (p-Nitrophenyl Phosphate, Disodium Salt) (Thermo Fisher Scientific) substrates for alkaline phosphatase and read at λ405 nm on Synergy H1 (BioTek Instruments).
Chodisetti S.B., Fike A.J., Domeier P.P., Schell S.L., Mockus T.E., Choi N.M., Corradetti C., Hou B., Atkins H.M., Caricchio R., Decker T., Lukacher A.E., Olsen N, & Rahman Z.S. (2020). Serine phosphorylation of the STAT1 transactivation domain promotes autoreactive B cell and systemic autoimmunity development. Journal of immunology (Baltimore, Md. : 1950), 204(10), 2641-2650.
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9

ELISpot Assay for Antibody-Producing Cells

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
ELISpot assays were performed as described (Wong et al., 2012 (link)). Briefly, splenocytes in RPMI containing 10% fetal bovine serum were plated at a concentration of 1 × 105 cells/well onto anti-IgM-, anti-IgG-, salmon sperm dsDNA- (Invitrogen, Grand Island, NY), calf thymus histone- (Sigma Aldrich, St. Louis, MO), or nucleosome-coated plates (Millipore, Bedford, MA). Serially diluted (1:2) cells were incubated for 6 hr at 37°C. IgM-producing AFCs were detected using biotinylated anti-mouse IgM (Jackson ImmunoResearch, West Grove, PA) and streptavidin (SA)-alkaline phosphatase (Vector Laboratories, Burlingame, CA). IgG-producing AFCs were detected using alkaline phosphatase-conjugated anti-mouse IgG (Molecular Probes, Grand Island, NY). dsDNA-, histone-, and nucleosome-specific AFCs were detected by biotinylated anti-kappa Ab (Invitrogen, Grand Island, NY) and SA-alkaline phosphatase or alkaline phosphatase-conjugated anti-mouse IgG (Molecular Probes, Grand Island, NY). Plates were developed using the Vector Blue Alkaline phosphatase Substrate Kit III (Vector Laboratories, Burlingame, CA). ELISpots were enumerated using a computerized ELISpot plate imaging and analysis system (Cellular Technology, Shaker Heights, OH).
Domeier P.P., Chodisetti S.B., Schell S.L., Kawasawa Y.I., Fasnacht M.J., Soni C, & Rahman Z.S. (2018). B-Cell-Intrinsic Type 1 Interferon Signaling Is Crucial for Loss of Tolerance and the Development of Autoreactive B Cells. Cell reports, 24(2), 406-418.
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10

Quantification of serum IgM/IgG and autoantibodies

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Total serum IgM/IgG or IgG from in vitro B cell culture supernatants was measured using standard ELISA protocols. Serum Ab titers were quantitated as described (29 ). Briefly, ELISA plates were coated with anti-IgM or anti-IgG capture antibodies (from Invitrogen, Grand Island, NY) and detected using biotinylated anti-mouse IgM (Jackson Immunoresearch, West Grove, PA) or alkaline-phosphatase conjugated anti-mouse IgG (Molecular Probes, Grand Island, NY). Total IgG autoAb titers were measured in ELISA plates coated with dsDNA, histone, nucleosome, Sm/RNP or cardiolipin and detected with biotinylated anti-kappa Ab (Invitrogen, Grand Island, NY). IgG subtype-specific autoAb titers were detected by biotinylated-IgG1, biotinylated IgG2b, and AP-IgG2c Abs (Southern Biotech, Birmingham, AL). Biotinylated antibodies were detected by streptavidin (SA)-alkaline phosphatase (Vector Laboratories, Burlingame, CA). The plates were developed by the PNPP (p-Nitrophenyl Phosphate, Disodium Salt) (Thermo Fisher Scientific, Rockford, IL) substrates for alkaline phosphatase.
Soni C., Wong E.B., Domeier P.P., Khan T.N., Satoh T., Akira S, & Rahman Z.S. (2014). B cell-intrinsic TLR7 signaling is essential for the development of spontaneous germinal centers. Journal of immunology (Baltimore, Md. : 1950), 193(9), 4400-4414.
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