Flow Cytometry FAQs

Antibody isotypes/subclasses are categorized based on the heavy chain structure of immunoglobulins (Ig). In mammals: 5 main Ig classes: IgA (α), IgD (δ), IgE (ε), IgG (γ), and IgM (μ). Human IgG subclasses: IgG1, IgG2, IgG3, IgG4. Mouse IgG subclasses: IgG1, IgG2a, IgG2b, IgG3. In flow cytometry: IgG subclasses (e.g., anti-CD3, CD4, CD8 for T cells; CD19, CD20 for B cells) are commonly used to detect surface antigens.

DAPI Working Solution: Add 4 μL of DAPI Reagent (25 μg/mL) to 96 μL of PBS and mix thoroughly. Prepare immediately before use. Tissue Sample Protocol: 1. After removing moisture with absorbent paper, add the DAPI working solution (cover the sample, and ensure the sample does not dry out during incubation). Incubate at room temperature in the dark for 5 minutes. 2. Immerse the sample in PBS and rinse 4 times, 5 minutes each time. 3. Remove excess liquid with absorbent paper, and mount the sample with an antifade mounting medium (self-prepared).

CD206 is a transmembrane protein expressed on both the cell surface and intracellularly. Our anti-mouse CD206 flow cytometry antibody can bind to CD206 on both the cell surface and intracellular regions. Therefore, it can be detected with or without permeabilization, depending on your experimental design. Generally, permeabilization is recommended for detection. If you choose to perform permeabilization, you can refer to the following experimental protocol: https://www.elabscience.cn/resource-flow_cytometry-detail-249

In most cases, the markers for naive CD4+ T cells are CD4+CD44-CD62L+ in mice and CD4+CD45RA+CCR7+ in humans. It is recommended to consult relevant literature to confirm the final detection indicators.

Yes, it is possible. However, it is important to note that the recommended antibody dosage may vary between brands, so the corresponding instructions for use should be followed accordingly. Additionally, for isotype controls, it is advisable to select products from the same brand as the primary antibody.

DAPI is not species-specific.

Since the sorting buffer contains serum, prolonged storage increases the risk of bacterial contamination. Therefore, it is recommended to prepare it fresh and use it immediately, with the storage time not exceeding one week.

No, this is not recommended. When anticoagulated whole blood is centrifuged, the upper layer is plasma, and the lower layer consists of a "blood cell pellet" (containing red blood cells, granulocytes, platelets, and a small number of white blood cells). After removing the plasma, the remaining sample is predominantly composed of red blood cells and granulocytes, with only a minimal proportion of PBMCs. If you attempt to stain the "blood cell pellet after plasma removal" directly, the excessive red blood cells, high background interference, and low PBMC ratio will result in unreliable data and may even clog the flow cytometer.

The cell yield from magnetic bead sorting (negative selection) varies significantly due to differences in samples. We cannot guarantee a specific value or range, but we can ensure that the purity of cells sorted using our magnetic bead sorting kit is consistently above 90%.

The primary purpose of red blood cell lysis buffer is to remove red blood cells from samples, while tissue lysis buffer is a reagent used to extract proteins, nucleic acids, or other biomolecules from tissues or cells.

For B cell subset analysis, it is sufficient to detect membrane-bound IgG, IgD, IgM, and IgA. Intracellular Ig detection (e.g., cytoplasmic IgG or IgA) requires fixation and permeabilization of the cell membrane.