Universal Enhanced IHC Secondary Antibody PLUS (HRP-conjugated)

货号MYE07

规格Solution A 60 ml + Solution B 60 ml (MYE07-1), Solution A 500 ml + Solution B 500 ml (MYE07-2), Solution A 1000 ml + Solution B 1000 ml (MYE07-4)

Secondary antibodies are core universal reagents in immunoassay systems. They enable indirect detection of target antigens by binding primary antibodies specifically and can significantly amplify detection signals to improve assay sensitivity.

Universal Enhanced IHC Secondary Antibody PLUS contains two reagents: enhancer (Solution A) and HRP-conjugated secondary antibody (Solution B). After the primary antibody binds the target antigen specifically, the enhancer creates numerous additional binding sites around the primary antibody, greatly increasing subsequent binding of HRP-conjugated secondary antibody. This technology preserves the compact structure and high affinity of the secondary antibody while concentrating far more HRP at each antigen site than conventional systems, achieving multi-level signal amplification for accurate detection of low-abundance targets.

Furthermore, this product eliminates the traditional biotin–streptavidin amplification pathway, avoiding false positives and nonspecific background staining caused by endogenous biotin in tissues, and thereby significantly improving contrast between positive signals and background.

    CD21 (MY149) shows strong membranous brown staining in most follicular dendritic cells in germinal centers of the tonsil, and moderate brown-yellow staining in mantle zone B cells. T cells and squamous epithelial cells show no staining.
    CD45 (MY221) shows moderate to strong cytoplasmic/membranous brown staining in almost all B cells and T cells in the tonsil.
    CDX-2 (MY235) shows strong nuclear/cytoplasmic brown staining in almost all tumor cells in colorectal cancer.
    Cytokeratin 5/6 (MY024) shows strong cytoplasmic brown staining in all squamous epithelial cells in the tonsil. Lymphocytes show no staining.
    Cytokeratin 7 (MY019) shows strong cytoplasmic brown staining in tumor cells and alveolar epithelial cells in lung adenocarcinoma. Lymphocytes, blood vessels, and fibroblasts show no staining.
    Cytokeratin 8 (MY061) shows moderate to strong predominantly membranous brown staining in hepatocytes in the liver, and strong cytoplasmic brown staining in bile ducts.
    Cytokeratin 20 (MY013) shows obvious cytoplasmic brown-yellow staining in tumor cells in colorectal cancer.
    E-cadherin (MY109) shows strong membranous brown staining in ductal epithelial cells in the breast.
    Estrogen Receptor α (MY007) shows moderate to strong nuclear brown staining in ductal epithelial cells in breast cancer. Tumor cells, myoepithelial cells, and stromal cells show no staining.
    Ki-67 (MY004) shows obvious moderate to strong nuclear brown staining in most lymphocytes in germinal centers of the tonsil.
    MSH2 (MY044) shows nuclear staining in glandular epithelial and stromal cells in the appendix, weak to moderate nuclear staining in mantle zone B cells and T cells in lymphoid nodules, and moderate to strong nuclear staining in B cells in germinal centers.
    MUC2 (MY054) shows obvious cytoplasmic brown staining in tumor cells in colorectal cancer.
    Napsin A (MY177) shows strong cytoplasmic brown staining in tumor cells in lung adenocarcinoma.
    p53 (MY081) shows strong nuclear staining in the vast majority of tumor cells in colorectal cancer.
    p57Kip2 (MY047) shows moderate to strong nuclear brown staining in trophoblast cells in the placenta. Syncytiotrophoblast cells show no staining.
    P63 (MY021) shows moderate to strong nuclear brown staining in squamous epithelial cells in the tonsil, with scattered lymphocytes showing weak to moderate nuclear brown-yellow staining.
    Progesterone Receptor (MY025) shows strong nuclear brown staining in tumor cells in breast cancer.
    S100 (MY072) shows moderate to strong nuclear/cytoplasmic brown staining in ganglion cells and axons in the appendix. Smooth muscle and glandular epithelial cells show no staining.
    TTF-1 (MY103) shows strong nuclear staining in tumor cells in lung adenocarcinoma.
    1. Soak tissue sections in two changes of fresh xylene for 10 min each, then in absolute ethanol, 95% ethanol, and 85% ethanol for 5 min each. Rinse gently in distilled water for 1 min.
    2. Place sections in Tris-EDTA (pH 9.0) antigen retrieval solution in a pressure cooker; heat until steaming and time for 2 min. After timing, allow to cool naturally for 5 min, then cool rapidly with water. Rinse gently in distilled water twice for 3 min each.
    3. Incubate sections in peroxidase blocking reagent (3% H2O2) for 10 min. Rinse gently in distilled water twice for 3 min each.
    4. Blot excess liquid around the tissue; circle the tissue edge 2–3 mm away with an IHC pen. Rinse gently in distilled water twice for 3 min each, then immerse in PBS buffer.
    5. Apply sufficient primary antibody working solution to cover the tissue completely. Incubate at room temperature in a humid chamber for 30–60 min. Wash with PBS 2–3 times and rinse gently twice for 3 min each.
    6. Apply sufficient secondary antibody Solution A to cover the tissue completely. Incubate at room temperature in a humid chamber for 10 min. Wash with PBS 2–3 times and rinse gently twice for 3 min each.
    7. Apply sufficient secondary antibody Solution B to cover the tissue completely. Incubate at room temperature in a humid chamber for 10 min. Wash with PBS 2–3 times and rinse gently twice for 3 min each.
    8. Apply sufficient DAB working solution to cover the tissue completely. Incubate at room temperature in a humid chamber for 5 min. Stop color development with distilled water.
    9. Counterstain in hematoxylin for 8–10 s. Rinse thoroughly with distilled water. Bluing in 1% lithium carbonate for 8–15 s, then rinse with distilled water.
    10. Dehydrate through 85% ethanol, 95% ethanol, and absolute ethanol for 3 min each, then in one change of fresh xylene for 5–10 min. Mount with neutral resin and examine under a microscope.
    Type
    Secondary antibody
    Form
    Liquid
    Shipping
    Biological refrigerated ice packs
    Storage conditions
    2–8°C
    Shelf life
    18 months
    Aliquoting
    Aliquoting for storage is recommended
    Storage notes
    Store protected from light and heat; avoid repeated freeze–thaw cycles