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Human iga elisa quantitation set

Manufactured by Fortis Life Sciences
Sourced in United States

The Human IgA ELISA Quantitation Set is a laboratory product designed to quantitate human immunoglobulin A (IgA) levels in biological samples using the enzyme-linked immunosorbent assay (ELISA) technique. The set includes the necessary components to perform the IgA ELISA, such as pre-coated microplates, standards, and reagents.

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5 protocols using human iga elisa quantitation set

1

Quantification of Human Milk Immunomodulators

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Human milk levels of TGF-β1, TGF-β2, and IgA levels were measured via commercial enzyme-linked immunosorbent assay (ELISA) kits. TGF-β1 and TGF-β2 levels were measured with a Human TGF-beta 1 DuoSet and a Human TGF-beta 2 DuoSet, respectively (R&D Systems, Minneapolis, MN, USA). IgA levels were measured with a Human IgA ELISA Quantitation Set (Bethyl Laboratories Inc., Montgomery, AL, USA). All ELISAs were conducted according to the respective manufacturers' instructions. The detection limits were 31.2 pg/mL for TGF-β1 and TGF-β2, and 7.8 ng/mL for IgA.
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2

T Cell Subset Interactions Modulate Antibody Production

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Each CD4+ T cell subset (1 × 105 cells/well) was cocultured with B cells (1 × 105 cells/well) and TFH cells (5 × 104 cells/well) in round-bottomed 96-well plates. Recombinant staphylococcal enterotoxin B (SEB) (2 μg/ml; Toxin Technology, Sarasota, FL, USA) was added to stimulate B cells. Blocking antihuman IL-10 mAb (25209; R&D Systems), blocking antihuman Fas ligand mAb (100410; R&D Systems), or mouse immunoglobulin G1κ (IgG1κ) isotype control (eBioscience) was added at a concentration of 10 μg/ml on the first day of the coculture in the same experimental system as above. To assess cell-cell contact, a coculture experiment was performed using a Transwell plate (HTS Transwell-96 System [3381]; Corning Life Sciences, Acton, MA, USA) with the following cells: B cells, 3 × 105 cells/well; TFH cells, 2 × 105 cells/well; and CD4+CD25LAG3+CD45RA T cells, 1 × 105 cells/well. The concentrations of IgG, IgA, and IgM in the 12-day culture supernatant were measured by ELISA (Human IgG ELISA Quantitation Set, Human IgA ELISA Quantitation Set, and Human IgM ELISA Quantitation Set; Bethyl Laboratories, Montgomery, TX, USA).
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3

B cell and PBMC ELISA Quantitation

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B cells and PBMCs were cultured at 3x105/well in 96 well plates. ELISA was performed using Human IgG ELISA Quantitation Set, Human IgA ELISA Quantitation Set, and Human IgM ELISA Quantitation Set (Bethyl Laboratories).
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4

Infant Gut Immunity Development

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Blood and stools of the subjects at birth and at one, two, and six months of age were collected. The collected blood samples were immediately tested at a laboratory in Shiga University of Medical Science Hospital to measure the serum IgG concentrations using the nephelometry method (BM6070, JEOL Ltd., Akishima, Japan). Serum IgG levels were calculated as the ratio of serum IgG concentrations at each month (one, two, and six months) to those at birth. The collected stool samples were immediately frozen at −20 °C. Then, the stool samples were mixed with nine volumes of phosphate-buffered saline and homogenized. The homogenates were centrifuged at 20,000× g for 15 min at 4 °C and the supernatant were used for quantification of SIgA. A commercial ELISA kit (Human IgA ELISA Quantitation Set, Bethyl Laboratories, Inc., Montgomery, TX, USA) was used to measure SIgA concentrations in the stool according by the manufacture’s instruction. Stool samples that were too small to be measured (<2.5 mg) were excluded. The technicians who measured the serum IgG and stool SIgA levels were blinded to the subjects’ groups.
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5

Quantification of Fecal and Breast Milk IgA

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The quantification of faecal IgA was performed as described previously [4 ]. Breast milk was centrifuged (9,100 × g, 10 min,
4°C), and the whey in it was used for the quantification of IgA using a commercial ELISA
kit (Human IgA ELISA Quantitation Set, Bethyl Laboratories, Inc., Montgomery, TX,
USA).
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