Anti-TLR4抗体[76B357.1] (ab22048)

Immunohistochemical analysis of paraffin-embedded Human skin tissue labeling TLR4 with ab22048 at 5 ug/ml. The section was blocked using blocking solution (1% BSA in PBS) for 1 hour at room temperature and incubated with ab22048 overnight at 4°C after removing blocking solution.

Antigen retrieval: Bring slides to a boil in 10 mM sodium citrate buffer (pH 6.0) then maintain at a sub-boiling temperature for 10 minutes. Cool slides on bench-top for 30 minutes (keep slides in the sodium citrate buffer all the time).

ab22048 staining TLR4 in Human stomach tissue sections by Immunohistochemistry (IHC-P - formaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 5% serum for 1 hour; antigen retrieval was by heat mediation in citrate buffer (10mM, pH 6) (ab64236). Samples were incubated with primary antibody (1/100) for 1 hour at 23°C. An undiluted HRP-conjugated goat anti-mouse IgG polyclonal was used as the secondary antibody.

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Overlay histogram showing Jurkat cells (ab7899) stained with ab22048 (red line). The cells were fixed with methanol (5 min) and incubated in 1x PBS / 10% normal goat serum (ab7481) / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab22048, 1µg/1x10⁶ cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in Jurkat cells fixed with 4% paraformaldehyde (10 min) used under the same conditions.

Please note that Abcam do not have data for use of this antibody on non-fixed cells. We welcome any customer feedback.

Representative examples of immunohistochemical staining of TLR4 in colorectal carcinoma tissues (original magnification 100×).

Paraffin-embedded human normal colorectal tissue (A) and colorectal carcinoma tissue (D) stained for TLR4 with ab22048 in immunohistochemical analysis. Counter stained with hematoxylin.

No staining is observed in normal colorectal tissue (A).

(From Figure 4A and 4D of Xu et al)

ab22048 (5 ug/ml) staining TLR4 in Human skin tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Membrane-cytoplasmic immunopositivity of TLR4 was primarily observed in the pigmented basel cells and the adjacent keratinocytes in the epidermal layer.

ab22048 at 5 µg/ml staining TLR4 in Human colon tissue sections by Immunohistochemistry (Formalin/ PFA-fixed paraffin-embedded tissue sections). Left hand image shows staining with an isotype control antibody whilst right one shows staining with ab22048.

ab22048 staining TLR4 in Rat's salivary gland tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). ab22048 at a dilution of 1:100 generated a membrane-cytoplasmic staining in the tissue with stronger signal in ductal epithelial cells.