This is a knockout-examined antibody summary, based on the publication "Specific detection of interferon regulatory factor 5 (IRF5): A case of antibody inequality", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

Figure 1. Immunoblot analysis of endogenous IRF5 protein expression in primary splenocytes from Irf5+/+ (WT) and Irf5−/− (KO) mice. β-actin levels and apparent molecular weight standards are shown as loading controls and for size comparison, respectively. Arrows indicate detection of an appropriately sized band(s) corresponding to murine IRF5. From [1].

Mouse monoclonal IgG1,kappa

Company: Abcam

Antibody: IRF5

Catalog number: ab33478

Summary: Mouse monoclonal IgG1,kappa against recombinant human IRF-5 (176-240 aa, isoform D). Reacts with human and mouse. Suitable for western blot, immunoprecipitation, immunocytochemistry, immunohistochemistry (paraffin), flow cytometry and ELISA.

Western blot

Primary splenocytes from Irf5+/+ (WT) and Irf5−/− (KO) mice.

5% BSA in TBS/0.05% Tween 20 for 1 h.

Overnight at 4 °C.

Horseradish peroxidase conjugated secondary antibodies (Cell Signaling: α-mouse #7076S).

Clarity™ ECL Western Blotting Substrate (Bio-Rad Laboratories) and chemiluminescence detected with a ChemiDoc™ MP Imaging System (Bio-Rad Laboratories).

Ab33478 (Abcam) detected a band in WT cells that corresponded to the size of IRF5 and was lacking in KO cells, yet numerous other non-specific bands were detected as well, indicating lack of specificity. Moreover, it was unable to IP IRF5.

IRF5 antibodies PT10547-1-AP (Proteintech), IC8447P (R&D), and SC98651 (Santa Cruz) were unable to detect differences in IRF5 expression between WT and KO splenocytes.

The same clone (10T1) is sold as Bio-Rad MCA2701; Santa Cruz Biotechnology sc-56714; LifeSpan Biosciences LS-C93755; Abnova MAB1068; Abcam ab33478; MyBioSource MBS214462, MBS200023; Antibodies-online ABIN121152.