TLR2 Recombinant Rabbit Monoclonal Antibody [JM22-41]

Western blot analysis of TLR2 on different lysates with Rabbit anti-TLR2 antibody (ET1705-92) at 1/1,000 dilution.

Lane 1: THP-1 cell lysate
Lane 2: HL-60 cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 90 kDa
Observed band size: 80-90 kDa

Exposure time: 30 seconds; ECL: K1801;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1705-92) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

☑ Knockout (KO)

All lanes: Western blot analysis of TLR2 with anti-TLR2 antibody [JM22-41] (ET1705-92) at 1/500 dilution.

Lane 1: Wild-type THP-1 whole cell lysate.
Lane 2: TLR2 knockout THP-1 whole cell lysate.

ET1705-92 was shown to specifically react with TLR2 in wild-type THP-1 cells. No band was observed when TLR2 knockout samples were tested. Wild-type and TLR2 knockout samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary Anti-TLR2 antibody (ET1705-92, 1/500) and Anti-β-actin antibody (R1207-1, 1/1,000) were used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG H&L (HRP) Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.

Cell lysate was provided by Ubigene Biosciences (Ubigene Biosciences Co., Ltd., Guangzhou, China).

Immunocytochemistry analysis of THP-1 cells labeling TLR2 with Rabbit anti-TLR2 antibody (ET1705-92) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-TLR2 antibody (ET1705-92) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

Flow cytometric analysis of TLR2 was done on THP-1 cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1705-92, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).