Tissue sections were deparaffinized in xylene and rehydrated with decreasing concentrations of alcohol. Subsequently, endogenous peroxide was blocked with hydrogen peroxide, and antigen retrieval was achieved by treating sections with 0.01 m sodium citrate, pH 6.0 for 1 min in a pressure cooker. After blocking with universal blocking solution (ZYMED Laboratories, San Francisco, CA, USA), tissue sections were stained with the designated primary antibodies. A Vectastain Elite ABC Peroxidase kit or Universal ImmPRESS kit (Vector Laboratories, Burlingame, CA, USA) were used for secondary antibodies, and visualization was performed using 3-amino-9-ethylcarbazole as a substrate (ZYMED Laboratories, San Francisco, CA, USA). Sections were then stained with hematoxylin for counterstaining and mounted using VectaMount AQ Aqueous Mounting Medium (Cat-No. H-5501; Vector Laboratories). Myeloperoxidase-positive (Abcam), F4/80-positive (Santa Cruz, 377009), and LY6C-positive cells (Abcam, ab15627) were counted in stained sections in six randomly chosen fields (×200), and bars are indicated standard errors of the means.
Universal immpress kit
Manufactured by Vector Laboratories
Sourced in United States
The Universal ImmPRESS kit is a versatile tool for immunohistochemistry and immunocytochemistry applications. It provides a polymer-based detection system that can be used with a wide range of primary antibodies, both polyclonal and monoclonal, against various species. The kit is designed to deliver a consistent and reliable signal amplification, enabling sensitive and efficient detection of target antigens in tissue sections or cell preparations.
Automatically generated - may contain errors
Lab products found in correlation
Vectamount aq aqueous mounting medium, by Vector Laboratories (1 mentions)
Vectastain elite abc peroxidase kit, by Vector Laboratories (1 mentions)
3 amino 9 ethylcarbazole, by Zymo Research (1 mentions)
Haematoxylin eosin, by Leica Biosystems (1 mentions)
Anti mpo, by Abcam (1 mentions)
Goat anti mouse il 1α, by R&D Systems (1 mentions)
Cyanine3 (cy3), by Jackson ImmunoResearch (1 mentions)
3 protocols using universal immpress kit
1
Immunohistochemical Analysis of Immune Cells
2
Paraffin-Embedded Tissue Immunostaining
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Tissue samples were embedded in paraffin. Paraffined tissues were cut into 4-micron microtome sections. Samples were stained with HAEMATOXYLIN/Eosin (LE-3801582E, LE-3801601E Leica biosystems) according to a standard protocol. Immunostaining was performed as previously described [24 (link)], using indicated first antibodies (Supplemental Table S3 ) and the Universal ImmPRESS kit (Vector Laboratories, Burlingame, CA, USA) as secondary antibodies.
The visualization was performed using the 3 amino-9-ethylcarbazole (AEC) substrate kit (SK-4205, ImmPACT, West Hills, CA, USA) and counterstained with hematoxylin. Sections were examined under a widefield microscope.
The visualization was performed using the 3 amino-9-ethylcarbazole (AEC) substrate kit (SK-4205, ImmPACT, West Hills, CA, USA) and counterstained with hematoxylin. Sections were examined under a widefield microscope.
3
Immunohistochemical and Immunofluorescence Analysis of Inflammatory Markers
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The following primary antibodies were used: rabbit polyclonal anti-MPO (Abcam, Inc., Cambridge, United Kingdom) and polyclonal rabbit-anti-mouse IL-17 (Santa Cruz Biotechnology, Inc.).
The Universal ImmPRESS kit (Vector Laboratories, Burlingame, CA) was used for secondary staining for immunohistochemistry, and visualization was performed using 3-amino-9-ethylcarbazole (AEC) as a substrate (Zymed Laboratories, San Francisco, CA). The numbers of IL-17- or MPO-positive cells were counted in stained sections in six randomly chosen fields (magnification, ×400), and the results are presented as averages. Goblet cells were stained with the commercial alcian blue kit (Bio-Optica, Milan, Italy).
For immunofluorescence studies, we used goat anti-mouse IL-1α (R&D, Minneapolis, MN, USA) and rabbit polyclonal anti-MUC2 (antimucin) antibodies (Abcam, Inc., Cambridge, England). Secondary antibodies were conjugated with Cy3 and Cy5 (Jackson ImmunoResearch, West Grove, PA). Sections were examined under a Zeiss laser-scanning confocal microscope.
The Universal ImmPRESS kit (Vector Laboratories, Burlingame, CA) was used for secondary staining for immunohistochemistry, and visualization was performed using 3-amino-9-ethylcarbazole (AEC) as a substrate (Zymed Laboratories, San Francisco, CA). The numbers of IL-17- or MPO-positive cells were counted in stained sections in six randomly chosen fields (magnification, ×400), and the results are presented as averages. Goblet cells were stained with the commercial alcian blue kit (Bio-Optica, Milan, Italy).
For immunofluorescence studies, we used goat anti-mouse IL-1α (R&D, Minneapolis, MN, USA) and rabbit polyclonal anti-MUC2 (antimucin) antibodies (Abcam, Inc., Cambridge, England). Secondary antibodies were conjugated with Cy3 and Cy5 (Jackson ImmunoResearch, West Grove, PA). Sections were examined under a Zeiss laser-scanning confocal microscope.
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