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Our Abpromise guarantee covers the use of ab13915 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||Use a concentration of 5 - 10 µg/ml. Permeabilization of samples might be necessary and overnight incubation with the ab13915 is recommended.|
|WB||Use a concentration of 1 - 3 µg/ml. Detects a band of approximately 130 kDa (predicted molecular weight: 108 kDa). TLR3 is a single-pass type I membrane protein. We recomend not to boil the samples, instead heat lysates at 60-70C for 10 minutes.|
|Flow Cyt||Use 2-5µg for 106 cells. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.|
|IP||Use 2µg for 106 cells.|
Immunohistochemical analysis of formalin fixed paraffin embedded tissue section of mouse intestine using ab13915 at 1/500 dilution with HRP-DAB detection and hematoxylin counterstaining. The image shows a punctate staining of the ER and endosomes in a subset of cells in Peyer's patches (organized lymphoid nodules) in the tested section.
Flow cytometry analysis of human B cells using ab13915 at 1µg/106 cells followed by FITC conjugated secondary antibody (Red). Green shows isotype control.
Western blot analysis of TLR3 in lysates from untransfected 293 cells (lane A), 293 cells transfected with human TLR3 cDNA (lane B), and 20
Immunohistochemical analysis of formalin fixed paraffin embedded tissue section of mouse colon using ab13915 at 1/500 dilution with HRP-DAB detection and hematoxylin counterstaining. Intense signal was found in subset of cells at the bases of the crypts.
ab13915 at 2µg / 1x106 cells staining intracellular TLR3 in human monocytes by Flow Cytometery. A PE-conjugated goat polyclonal to mouse IgG1 was used as secondary. The shaded histogram repersents monocytes without antibody; green repersents isotype control and red histogram shows staining with ab13915.
ab13915 at 5ug/ml staining TLR3 in mouse spleen tissue section by Immunohistochemistry (Formalin/ PFA fixed paraffin-embedded sections). The left image repersents staining with isotype control and right one repersents staining with ab13915.
ab13915 staining TLR3 in SW480, SW620 (colon cancer) and FaDu, Detroit 562 obtained from human cells by Flow Cytometry. Samples were prepared by trypsinizining and washing in PBS and centrifugation in a buffer containing 2% FBS, 0.1% Na-azide and PBS. Cells were fixed in paraformaldehyde and permeabilized in 0.2% Tween. Gating and analysis was performed by WinMDI 2.9 software. The primary antibody was diluted to 1 µg/cells (PBS, 2% FBS and 0.1% Na-azide) and incubated with sample for 30 minutes at 4°C. An FITC conjugated rabbit polyclonal to mouse IgG, diluted to 1/40, was used as secondary. Image represents FACS analysis of TLR3 protein expression in SW480 (A), SW620 (B), FaDu (C) and Detroit 562 (D) cell lines. Cell lines were permeabilized and immunostained with TLR3-specific antibody (thick black l