BeaverBeads® Mouse Pan B Cell Isolation Kit (Negative Selection)
Product Introduction
BeaverBeads® Mouse Pan B Cell Isolation Kit isolates total Bcells from mouse spleen, lymph node, bonemarrow cells or peritoneal lavage fluid via negativeselection methodology. Biotinconjugated monoclonal antibodies are used to label nontarget cells (nonBcells), which are then depleted by streptavidincoated magnetic beads to achieve purification of mouse total Bcells.
Product Features
High Purity: The purity of isolated mouse B cells can reach up to 98.8%.
High Viability: Well-functioning mouse total B cells without antibody labeling can be obtained.
Rapid & Simple Workflow: No separation column required. Isolation can be completed within 25 minutes using a magnetic separator.
Performance Data
1. Cell Purity Analysis
Total Bcells were isolated from bonemarrow cells and splenocytes of C57BL/6 mice using the BeaverBeads® Mouse Pan BCell Isolation Kit (Negative Selection). Cells pre and postisolation were stained with FITC antimouse CD3 antibody (Clone: 1452C11) and APC antimouse CD19 antibody (Clone: 6D5), followed by flow cytometric analysis. As shown in Figure 1, Bcell purity was 63.8 % before isolation and reached 98.8 % after isolation for splenocytes; for bonemarrow cells, Bcell purity increased from 28.2 % to 91.1 % postisolation.
Figure 1. Flowcytometric analysis of Bcell purity pre and postisolation
2. CellPurity Analysis for Peritoneal LavageFluid Samples and Preservation & Identification of B1 Bcells
Purity analysis of isolated CD19⁺ Bcells
Total Bcells were isolated from peritoneal lavage fluid of healthy C57BL/6 mice with the BeaverBeads® Mouse Pan B Cell Isolation Kit (Negative Selection). Cells before and after isolation were stained with FITC antimouse CD3 antibody (Clone: 1452C11), PE antimouse CD11b antibody (Clone: M1/70) and APC antimouse CD19 antibody (Clone: 6D5) for flow cytometric measurement. As presented in Figure 2, Bcell purity in peritoneal lavage fluid rose from 30.9 % preisolation to 97.1 % postisolation, and the CD11bᵈⁱᵐ cell population was well preserved among sorted CD19⁺ cells.
Figure 2. Flowcytometric analysis of Bcell purity pre and postisolation from peritoneal lavage fluid
Preservation and identification of sorted B1 Bcells
Total Bcells were isolated from peritoneal lavage fluid of healthy C57BL/6 mice using the BeaverBeads® Mouse Pan B Cell Isolation Kit (Negative Selection). Pre and postisolation cells were stained with FITC antimouse B220 antibody (Clone: RA36B2), APC antimouse CD5 antibody (Clone: 537.3) and PE antimouse CD23 antibody (Clone: B3B4) for flow cytometry. As shown in Figure 3, cell populations of CD5⁻ and CD5⁺ were analysed within the CD23⁻ B220ᵈⁱᵐ gate (Gate R2). The proportion of CD23⁻ B220ᵈⁱᵐ CD5⁺ (B1) cells was 23.6 % before isolation and increased to 59.8 % after isolation.
Figure 3. Flowcytometric analysis of B1 Bcell purity pre and postisolation from peritoneal lavage fluid
3. Cell Viability Analysis
Total Bcells were isolated from splenocytes and bonemarrow cells of C57BL/6 mice using the BeaverBeads® Mouse Pan B Cell Isolation Kit (Negative Selection). Cells before and after isolation were stained with 7AAD and analysed by flow cytometry. As shown in Figure 4, splenocyte viability was 86.3 % preisolation and 89.9 % postisolation. Bonemarrowcell viability was 95.3 % before isolation and 96.7 % after isolation.
Figure 4. Flowcytometric detection of Bcell viability
4. Cellular Functional Assay
Total Bcells were isolated from splenocytes of C57BL/6 mice with the BeaverBeads® Mouse Pan BCell Isolation Kit (NegativeSelection). Purified Bcells were stimulated with LPS for 24 h, and surface activationmarker expression was assessed by flow cytometry. As illustrated in Figure 5, LPS stimulation markedly upregulated surface expression of CD69, CD80 and CD86 on Bcells.
Figure 5. Upregulation of surface activation markers on Bcells upon LPS stimulation
5. Performance Comparison with Competitor Kits
Total B cells purity and viability obtained with the BeaverBeads® Mouse Pan B Cell Isolation Kit (Negative Selection) were compared against those of a commercial competitor Pan B cell isolation kit.
(1) Total B cells were isolated from splenocytes, bonemarrow cells and peritoneal lavage fluid of C57BL/6 mice. Pre and postisolation cells were labelled with APC antimouse CD19 antibody (Clone: 6D5) for flowcytometry analysis. As shown in Figure 6, Bcell purity achieved by the BeaverBeads® kit exceeded that of the competitor control group across all sample types.
Figure 6. Comparison of mouse total Bcell isolation purity between BEAVER kit and competitor kit
(2) Total Bcells were isolated from splenocytes and bonemarrow cells using respectively the Mouse Pan BCell Isolation Kit (NegativeSelection) and the competitor kit, and cell viability was compared. As shown in Figure 7, the BeaverBeads® Mouse Pan BCell Isolation Kit (NegativeSelection) delivered superior Bcell viability for both splenocytes and bonemarrow cells relative to the competitor control.
Figure 7. Comparison of mouse Bcell isolation viability between BEAVER kit and competitor kit
