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Recombinant human TLR2 produced by the CHO/CD14 reporter cell line.
Ab9100 is a TLR2 function blocking antibody that is useful for studies on the role of TLR2 as a pattern recognition receptor in microbial products induced cytokine production by TLR2 bearing cells such as human pheriperal blood mononuclear cells.
Our Abpromise guarantee covers the use of ab9100 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use a concentration of 5 - 10 µg/ml.
Fix with PBS/2% formalin for 15 min at 4°C; permeabilize with acetone, 10 min, -20°C (see Flo et al J Leukoc Biol which used a conjugated form of the antibody).
|Flow Cyt||Use 2-3µg for 105 cells.
Untreated and 0.4 % formaldehyde fixed cells can be used; do not permeabilize the cells.
ab170191-Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.
|IP||Use at an assay dependent concentration.|
|Neutralising||Use at an assay dependent concentration. See Lien et al.|
Abcam recommends using non-reducing conditions.
|Functional Studies||Use at an assay dependent concentration.|
This blot was produced using a 4-12% Bis-Tris gel under the MOPS buffer system. The gel was ran at 200V for 60 minutes before being transferred onto a nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked with 2% BSA for 60 minutes before being incubated with ab9100 (anti-TLR2 antibody; 1 ug/mL) for 18 hours at 4°C. Antibody binding was detected using ab97040 (HRP-labelled goat anti-mouse IgG) at 1:50,000 dilutions for 1 hour at room temperature.Visualisation was conducted using ECL development solution ab133406.
This gel was run under reducing conditions resulting in weak signal. We recommend customers to use non-reducing conditions.
ab9100 staining TLR2 in human U937 cells by Immunocytochemistry. Cells were fixed with paraformaldehyde and blocking with 2% BSA was done for 30 minutes at 270C. Samples were incubated with primary antibody (1/50) for 1 hour at 27°C. An Alexa Fluor®568-conjugated goat polyclonal to mouse IgG (H&L) was used at dilution at 1/50 as secondary antibody.
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