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IMG-416E [Monoclonal Antibody to human TLR2 (Toll-like receptor 2)/CD282 Azide free ]
Does this antibody work for neutralization or blocking? If so, can you provide some form of starting protocol?
Answer :
Hi and thank you for your email. IMg-416, clone TL2.1 has been published for use in neutralization studies. The amount of antibody needed will depend on the model system used, and the amount of ligand to be blocked (as well as the number of cells). Researchers will need to titrate. However the following is provided as a guideline for TLR2 Neutralization of Cytokine Production:
Materials Needed
1. Human cells of interest 2. Culture Medium, such as RPMI or IMDM supplemented with 10% FBS 3. 24-well flat-bottom culture plate 4. IMGENEX anti-human TLR2, clone TL2.1 (Cat. No. IMG-416E)
Method
Prepare cells of interest at a concentration of 2x106 cells per ml in 10% FBS-RPMI/IMDM (plus 5% human serum).
Pre-incubate cells with 10-20 µg/ml of TL2.1 at room temperature for 30min.
Add the ligand and incubate at 37°C, 5% CO2 for 14 hours.
Harvest supernatants.
Assay for to cytokine levels. Normally this would be for IL-6 or TNF-a.
Could you recommend good antobodies of Toll-like receptor 4 and Toll-like receptor 2 for immunohistochemistry analysis?
Answer :
Thank you for your interest in the TLR antibodies. We have not evaluated TLR2 antibodies for immunohistochemistry. The applications developed for the antibodies are listed on Website: http://imgenex.com/search_result.php?txtSearch=tlr2 . A researcher would need to determine (validate) for themselves if the antibodies were suitable for IHC and then optimize in their own hands. You may want to click on the links for the data sheets to find out additional information about the TLR2 antibodies to determine if one may be suitable for your studies.
One of the TLR4 antibodies has been published for immunofluorescence. The citation is listed on the data sheet. Please follow the link to determine if the product may be suitable for your application: http://imgenex.com/antibody_details.php?catalog=IMG-417A . The known applications of the all of the TLR4 antibodies can also be found on the website by entering the search term "TLR4" on the home page.
Keyword(s):
TLR, TLR2, TLR4, IHC, IF, immunofluorescence
Question : 3
Does anti-TLR2 (IMG-416E) act as an antagonist antibody?
For how long does the antibody stay attached to the receptor once it is applied to the cell culture?
How long should I incubate the cell culture with the antiboty prior to adding any competative influence?
Answer :
As denoted on the data sheet, the Neutralization assays were first described by Flo et al, 2002 (see product citation 12 on the data sheet). There are additional citations for this antibody in the list denote as FA for functional assays. At Imgenex, we routinely quality control each batch of antibody by flow cytometry as shown on the data sheet. The antibody binds to the TLR2 receptor and acts an antagonist antibody, this feature was also characterized in the Flo et al, 2000 reference as denoted on the data sheet link. I am not aware of any studies which have been done to determine how long the antibodies stays attached to the receptor once it is in culture. With regards to incubation length, in the Flo et al 2000 reference, the antibody was added 30 min prior to adding the stimuli (in this case LPS or listeria). You may also want to look at the other product citations where the antibody was used for Functional assays to obtain additional information on the incubation topic.
