Six‐ to eight‐week‐old C57BL/6J mice were purchased from the Jackson Laboratory (Bar Harbor, Maine). Mouse experiments were approved by the MUSC IACUC. Pulmonary fibrosis was induced in mice following intratracheal administration of 1.2 mU/g bleomycin mixed with 10 µg rhIGFBP‐4 or vehicle in a volume of 50 µL. rhIGFBP‐4 or mutant IGFBP‐4 were administered to different mice on days 0, 3, and 6 post‐bleomycin for a total of three doses. On day 21, mice were euthanized and lungs were gently perfused with 1X PBS. The left lung was collected for hydroxyproline assay. The right lung was further perfused with 10% zinc formalin fixative (Polysciences Inc, Warrington, PA) and fixed in 10% zinc formalin for 48 hours prior to paraffin embedding and histological analysis.
Zinc formalin fixative
Manufactured by Polysciences
Sourced in United Kingdom
Zinc formalin fixative is a chemical solution used in laboratory settings for the preservation and fixation of biological samples. It is composed of a combination of zinc salts and formalin. The primary function of this fixative is to maintain the structural integrity of tissue samples, enabling effective preservation and subsequent analysis. This product is intended for use in research and diagnostic applications within controlled laboratory environments.
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Lab products found in correlation
3 protocols using zinc formalin fixative
1
Bleomycin-Induced Pulmonary Fibrosis in Mice
2
Immunohistochemical Analysis of IL-22BP and pSTAT3
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PP tissues were fixed with zinc formalin fixative (Polysciences) and embedded into paraffin. Paraffin sections of PPs were deparaffinized and treated with 0.3% H2O2 in PBS for 20 min at room temperature to quench an endogenous peroxidase activity and then sections were boiled with citrate buffer, pH 6.0, for 20 min to antigen retrieval of both IL-22BP and pSTAT3. The sections were incubated with 0.5% blocking buffer (PerkinElmer) in PBS for 30 min at room temperature and then with primary antibodies overnight at 4°C. The binding of anti–IL-22BP (AF1087; R&D Systems) and anti–phospho-STAT3 (Tyr705; Cell Signaling Technology) was followed by biotinylated secondary antibodies and visualized by a Tyramide Signal Amplification system (PerkinElmer). The sections were analyzed with an SP5 confocal laser microscope (Leica Microsystems).
3
Tumor PD-L1 Expression Analysis
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Tumor cells were subcutaneously implanted into C57BL/6 mice for tumor growth. Tumors were collected 3 weeks after implantation, fixed with Zinc Formalin Fixative (Polysciences Inc, Warrington, PA; #21516), and embedded in paraffin. Tumor sample paraffin sections were used for RNA in situ hybridization analysis by using the RNAscope 2.5 HD Assay – BROWN (Advanced Cell Diagnostics, Newark, CA) with probe targeting CD274 (PD-L1) (420501). The RNA in situ hybridization analysis was performed following standard procedures from the kit manual and published protocol32 (link) (RNAscope: a novel in situ RNA analysis platform for formalin-fixed, paraffin-embedded tissues).
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