This is a knockout-validated 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].

Rabbit monoclonal IgG

Company: Abcam

Antibody: IRF5

Catalog number: ab181553

Summary: Rabbit monoclonal IgG against a recombinant fragment within human IRF5 aa 1-200. Reacts with human, mouse and rat protein. Suitable for western blotting and immunoprecipitation, immunohistochemistry (paraffin), immunofluorescence and flow cytometry.

Figure 2. Representative images from IF (left panels) and IHC (right panels) analysis of IRF5 expression in spleen from Irf5+/+ (WT) and Irf5−/− (KO) mice. For IF, IRF5-positive staining is red; DAPI is blue. For IHC, IRF5-positive staining is brown; hematoxylin is blue; 2nd alone, staining with secondary antibody only. From [1].

Western blot | Immunofluorescence | Immunohistochemistry

WB: Primary splenocytes from Irf5+/+ (WT) and Irf5−/− (KO) mice. IF and IHC: spleen from Irf5+/+ (WT) and Irf5−/− (KO) mice.

WB: 5% BSA in TBS/0.05% Tween 20 for 1 h.

IF: 4% BSA for 1 h.

IHC: BloXall (Vector Laboratories: #SP-100) for 10 min at room temperature, Avidin/Biotin Blocking Kit (Vector Laboratories: #SP-2001), and 5% normal serum in 0.1% Triton X-100.

WB: overnight at 4 °C.

IF: 1:150 dilution overnight at 4 °C.

IHC: 1:100 dilution in serum blocking buffer overnight at 4 °C.

WB: Horseradish peroxidase conjugated secondary antibodies (Cell Signaling: α-rabbit #7074S, α-mouse #7076S; Santa Cruz: donkey α-goat #sc-2020).

IF: 1:1,000 dilution of secondary antibodies (Jackson ImmunoResearch: Cy3-anti-rabbit #711-165-152) for 1 h.

IHC: Biotin-conjugated secondary antibodies.

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

IF: Zeiss Axiovert 200 M System (Zeiss).

IHC: avidin-biotin complex (Vector Laboratories: VECTASTAIN Elite ABC Kit #PK-6101) followed by ImmPACT™ DAB Substrate Kit (Vector Laboratories: #SK-4105).

If the antibody described in this summary is a polyclonal antibody, since polyclonal antibodies are of limited quantity, please inquire the supplier whether any current polyclonal antibody with the same catalog number is exactly the same as the one described in this summary. Sometimes, different bleeds or different animals are used, usually with a different lot number. In such cases, the result in this summary may not apply to the new antibody with the same catalog number.