DSA were measured, as described previously (12 (link)). In brief, serum of recipient mice was obtained after rejection or at the end of follow-up. After heat inactivation, serum was incubated with donor‐ or recipient‐type thymocytes. Binding of IgG on thymocytes was measured by staining with anti–mouse IgG1‐FITC, IgG2a/2b‐FITC and IgG3-FITC (BD Biosciences) utilizing flow-cytometrical analysis on a BD CANTO II (BD Biosciences).
Anti mouse igg1 fitc
Manufactured by BD
Anti–mouse IgG1‐FITC is a fluorescently labeled secondary antibody that binds to mouse IgG1 antibodies. It can be used to detect and visualize mouse IgG1 proteins in various applications.
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Lab products found in correlation
2 protocols using anti mouse igg1 fitc
1
Flow Cytometric Measurement of Donor-Specific Antibodies
2
Donor-Specific Antibody Detection and T-Cell Proliferation Assay
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Donor-specific antibodies (DSAs) were measured by flow cytometry. Serum was obtained from recipients at the time of rejection or at 100 days after transplant. After heat inactivation, serum was incubated with donor-or recipient-type thymocytes. Binding of IgG was measured by staining with anti-mouse IgG1-FITC and IgG2a/2b-FITC (BD Biosciences). ). In brief, unseparated splenocytes (4 × 10 5 ) were incubated in triplicate with 4 × 10 5 irradiated (30 Gy) BALB/c (donor) cells, with C57BL/6 (recipient) stimulator cells or with medium for 72 h. Cells were pulsed with [3H]-thymidine (Amersham BioSciences, Little Chalfont, U.K.) for 18 h, and incorporated radioactivity was measured with a MicroBeta 1450 beta counter (Wallac; HVD Lifesciences, Vienna, Austria). Stimulation indices were calculated by dividing mean counts per minute of response against recipient or donor by mean background counts per minute.
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