Affinity Purification of Anti-Mouse COL7 IgG

Affinity purified rabbit anti-mouse COL7 IgG was produced as described (10 (link)). Briefly, rabbits were immunized with an immunodominant fragment of murine COL7. Total immune IgG was isolated from rabbit serum using protein G, followed by isolation of COL7-specific IgG using COL7 coupled to an Affi-Gel 10 matrix (Bio-Rad #1536099). COL7 affinity purified IgG was labeled with DyLight™ 594 NHS ester (Thermo Scientific #46412) according to manufactures instructions. Monoclonal FcγRIV blocking antibody 9E9 was kindly provided by Falk Nimmerjahn (9 (link)), the isotype control IgG1κ (clone A19-3) was purchased from BD Biosciences (#553968).

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Haeger S.C., Kridin K., Pieper M., Griewahn L., Nimmerjahn F., Zillikens D., König P., Ludwig R.J, & Hundt J.E. (2022). Therapeutic effects of Fc gamma RIV inhibition are mediated by selectively blocking immune complex-induced neutrophil activation in epidermolysis bullosa acquisita. Frontiers in Immunology, 13, 938306.

Publication 2022

DyLight™ 594 NHS ester

Affi gel 10 Anti igg Blocking antibody Clone Ester Isolation Isotype Mouse Murine Protein g Rabbit Rabbits Serum

Corresponding Organization : University of Lübeck

Other organizations : Bar-Ilan University, Friedrich-Alexander-Universität Erlangen-Nürnberg

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Variable analysis

independent variables

Immunization of rabbits with an immunodominant fragment of murine COL7

dependent variables

Isolation of total immune IgG from rabbit serum
Isolation of COL7-specific IgG using COL7 coupled to an Affi-Gel 10 matrix
Labeling of COL7 affinity purified IgG with DyLight™ 594 NHS ester

control variables

Protein G for isolation of total immune IgG from rabbit serum
Affi-Gel 10 matrix for isolation of COL7-specific IgG
Manufacturer's instructions for labeling COL7 affinity purified IgG with DyLight™ 594 NHS ester

positive controls

Monoclonal FcγRIV blocking antibody 9E9 provided by Falk Nimmerjahn

negative controls

Isotype control IgG1κ (clone A19-3) purchased from BD Biosciences

Annotations

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