Unstained sections were de-paraffinized with xylenes and alcohols prior to unmasking of antigens. Sections were washed in PBS containing triton X-100 and incubated in blocking solution for 1 hour, prior to incubation with primary antibodies for TLR-4, TGFβ1, pNFkB, and IkB (Santa Cruz Biotechnology (Dallas, TX) overnight. IHC-Tek antibody diluent (pH 7.4) purchased from IHC World (Woodstock, MD) was used for negative controls. The sections were incubated with Alexa Fluor fluorescent secondary antibodies (ThermoFisher Sci/Life Technologies, Grand Island, NY) and counterstained with DAPI. Images were captured at 20X magnification (scale bar=50 μm) using an Olympus BX53 microscope, DP72 digital camera, and CellSens imaging software attached to a Dell Precision T3500 computer (Olympus America, Inc. Center Valley, PA). Quantitative analysis of the stain intensity was conducted using the count and measure on ROI tool of the CellSens software.
Ihc tek antibody diluent ph 7.4
Manufactured by IHC World
IHC-Tek antibody diluent (pH 7.4) is a laboratory solution designed to dilute antibodies prior to their use in immunohistochemistry (IHC) applications. It provides a buffered environment with a pH of 7.4 to maintain the stability and activity of the antibodies.
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Lab products found in correlation
2 protocols using ihc tek antibody diluent ph 7.4
1
Quantitative Immunofluorescence Analysis of TLR-4, TGFβ1, pNFκB, and IκB
2
Quantitative Immunofluorescence Analysis of TLR-4, TGFβ1, pNFκB, and IκB
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Unstained sections were de-paraffinized with xylenes and alcohols prior to unmasking of antigens. Sections were washed in PBS containing triton X-100 and incubated in blocking solution for 1 hour, prior to incubation with primary antibodies for TLR-4, TGFβ1, pNFkB, and IkB (Santa Cruz Biotechnology (Dallas, TX) overnight. IHC-Tek antibody diluent (pH 7.4) purchased from IHC World (Woodstock, MD) was used for negative controls. The sections were incubated with Alexa Fluor fluorescent secondary antibodies (ThermoFisher Sci/Life Technologies, Grand Island, NY) and counterstained with DAPI. Images were captured at 20X magnification (scale bar=50 μm) using an Olympus BX53 microscope, DP72 digital camera, and CellSens imaging software attached to a Dell Precision T3500 computer (Olympus America, Inc. Center Valley, PA). Quantitative analysis of the stain intensity was conducted using the count and measure on ROI tool of the CellSens software.
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