Recombinant Anti-TLR9 antibody [EPR21735] - BSA and Azide free (ab232933)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab211012).

False colour image of Western blot: Anti-TLR9 antibody [EPR21735] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab211012 was shown to bind specifically to TLR9. A band was observed at 140 kDa in wild-type Raji cell lysates with no signal observed at this size in TLR9 knockout cell line ab280879 (knockout cell lysate ab282939). To generate this image, wild-type and TLR9 knockout Raji cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

Blocking/Dilution buffer: 5% NFDM/TBST.

The cleaved TLR9 (60 kDa) observed is consistent with the literature (PMID: 24582318).

Untreated THP1s have undetectable TLR9, as described in the literature (PMID: 20375564).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab211012).

TLR9 was immunoprecipitated from 0.35 mg of Raji (human Burkitt's lymphoma cell line) whole cell lysate with ab211012 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab211012 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

Lane 1: Raji whole cell lysate 10 µg (Input). 

Lane 2: ab211012 IP in Raji whole cell lysate. 

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab211012 in Raji whole cell lysate.

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 10 seconds.

The cleaved TLR9 (60 kDa) observed is consistent with the literature (PMID: 24582318).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab211012).