| Product Description | Immunostaining Permeabilization Buffer with Saponin can be used for permeabilization of cell samples, frozen or paraffin sections in in situ assays such as immunostaining to expose the detection targets such as antigens and nucleic acids, allowing for efficient target labeling by antibodies, probes, or other markers. This ready-to-use product can be used directly without any extra preparations.This product has strong permeability and is recommended for cell permeabilization in various routine immunofluorescence, immunohistochemistry, immunocytochemistry, and flow cytometry assays, etc. It is especially suitable for detection of cell membrane proteins. This product does not dissolve cell membrane nor affect the light scattering during flow cytometry assay. However, this product is not recommended for detection of nuclear and mitochondrial proteins. For low permeation requirements, Aladdin's Immunostaining Wash Buffer or Immunostaining Permeabilization Buffer with Triton X-100 can also be used. For higher permeation requirements, Enhanced Immunostaining Permeabilization Buffer can be used. For lectin detection, Immunostaining Permeabilization Buffer with Triton X-100 or Enhanced Immunostaining Permeabilization Buffer is recommended.Permeabilization of cell membranes is usually achieved by using organic solvents such as methanol and acetone, or by using detergents such as Triton X-100 and Saponin. Organic solvents can lyse cell membrane and nuclear membrane to fully expose target proteins in cell plasma and nucleus, and also denature proteins in the meantime as a fixative. The disadvantage is that membrane proteins are also dissolved and some proteins are denatured, which is unfavorable sometimes for downstream applications. Therefore, organic solvents are rarely used except for some rough assays. Triton X-100 is a commonly used permeabilizing reagent that can permeate cell and nuclear membranes by non-specific lysis of cell membranes, thus its disadvantage is that it is not conducive for detection of membrane proteins. However, after cross-linking and fixation with paraformaldehyde, a significant portion of membrane proteins will not be lysed by Triton X-100 and can be detected subsequently. Saponin specifically solubilizes cholesterol in cell membranes and thus enables selective perforation on cell membranes. Its advantage is that it is suitable for examining cell membrane proteins, especially signature proteins on cell membranes by flow cytometry. Its disadvantage is that it is poorly permeable to cells with low cholesterol content, with weaker permeability than Triton X-100 and organic solvents, and cannot penetrate nuclear and mitochondrial membranes that have very low cholesterol content. For the detection of lectin, permeabilization solutions containing non-specific detergents such as Triton X-100 are significantly more effective than those containing Saponin mainly.This product contains Saponin and other detergents and is prepared in PBS. Immunostaining assay results of target cell membrane proteins in samples treated with this product is basically the same or even superior to that obtained with conventional permeabilization solutions.
Precautions： This product is for R&D only. Not for drug, household, or other uses.For your safety and health, please wear a lab coat and disposable gloves during the operation.
Instructions for Use： 1. For slice sections, after fixation and washing, add 50-100μl of this product per sample, or permeabilize by complete submersion in this product in a staining jar. For cell samples, after fixation and washing, add 1ml of this product per well for six-well plates, or refer to this ratio for cells in other multi-well plates. For other samples, add sufficient Immunostaining Permeabilization Buffer to cover samples completely.2. Permeabilization can usually be completed by incubation at room temperature for 5-10 minutes. For samples that are more difficult to permeate or require full permeation particularly, incubate for 10-30 minutes at room temperature.
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