Freshly sorted cells or day-2-post culture cells were tested for antibody secretion by ELISPOT. Sorted cells were plated on goat anti-human IgM or goat anti-human IgG coated plates in medium alone, and incubated in 5% CO2 for 14-18 hours at 37° C. Spot-forming cells were detected by incubation with Alkaline Phosphatase-conjugated goat anti-human IgM or anti-human IgG (Southern Biotech), and developed with VECTOR Blue, Alkaline Phosphatase Substrate Kit III (Vector Laboratories). Spots in each well were counted using a C.T.L ImmunoSpot plate reader and counting software (Cellular Technology Limited).
Alkaline phosphatase conjugated goat anti human igm or anti human igg
Manufactured by Southern Biotech
Alkaline Phosphatase-conjugated goat anti-human IgM or anti-human IgG is a laboratory reagent used for the detection and quantification of human immunoglobulin M (IgM) or immunoglobulin G (IgG) in various immunoassay applications. The antibody is conjugated with the enzyme alkaline phosphatase, which can catalyze the conversion of colorless substrates into colored products, enabling the visualization and measurement of the target analyte.
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Lab products found in correlation
2 protocols using alkaline phosphatase conjugated goat anti human igm or anti human igg
1
Antibody Secretion ELISPOT Assay
2
Antibody-Secreting Cell Enumeration by ELISPOT
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ELISPOT assays were used to detect antibody-secreting cells. Sort-purified cells (1–10 x 103) were distributed on Multi-Screen-IP Plates (Millipore) precoated with goat anti–human IgM or goat anti–human IgG and then incubated in complete medium (RPMI medium supplemented with 10% FCS, 10 mM HEPES, 1 mM non-essential aminoacids, 2 mM L-glutamine, 1 mM pyruvate, and 50 μg/mL Penicillin/Streptomycin) for 18 h at 37°C and 5% CO2. Plates were then incubated with alkaline phosphatase-conjugated goat anti-human IgM or anti-human IgG (Southern Biotech), and developed with VECTOR Blue, Alkaline Phosphatase Substrate Kit III (Vector Laboratories). Immunoglobulin-secreting B cells were enumerated using C.T.L ImmunoSpot software (Cellular Technology Limited).
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