Preparation of IgG anti-SRBC allotypes

SRBC were obtained from Håtunalab AB (Håtunaholm, Sweden) and were stored in sterile Alsever's solution at 4°C. IgG anti-SRBC of the Ig^a and Ig^b allotypes were prepared as described (23 (link)). Briefly, BALB/c and C57BL/6 mice were immunized i.v. with 10% SRBC three times. Sera from blood obtained 4 weeks after the last immunization were run over a Protein-A Sepharose column (Amersham Pharmacia Biotech, Uppsala, Sweden). The eluted IgG fractions were dialyzed, concentrated and stored at −20° until use.

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Anania J.C., Westin A, & Heyman B. (2020). IgG Suppresses Antibody Responses to Sheep Red Blood Cells in Double Knock-Out Mice Lacking Complement Factor C3 and Activating Fcγ-Receptors. Frontiers in Immunology, 11, 1404.

Publication 2020

Protein-A Sepharose column

A protein Anti igg Blood sera C57bl mice Immunization Sepharose Sterile

Corresponding Organization :

Other organizations : Uppsala University

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Variable analysis

independent variables

Ig^a and Ig^b allotypes of IgG anti-SRBC

dependent variables

Not explicitly mentioned

control variables

Storage of SRBC in sterile Alsever's solution at 4°C
Immunization of BALB/c and C57BL/6 mice with 10% SRBC three times
Serum collection 4 weeks after the last immunization
Purification of IgG fractions using Protein-A Sepharose column
Dialysis and concentration of the eluted IgG fractions
Storage of the IgG fractions at -20°C until use

controls

Positive control: Immunization of BALB/c and C57BL/6 mice with 10% SRBC three times
Negative control: Not explicitly mentioned

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