Cells were incubated in 50 μL of FACS buffer containing a cocktail of antibodies for 30 min on ice prior to washing and analysis on a FACS Aria (BD). Antibodies included anti-IgM FITC (G20-127, BD), anti-CD19 BUV395 (SJ25C1, BD), anti-CD3 BV711 (UCHT1, BD), anti-CD14 BV711 (M0P-9, BD), anti-CD16 BV711 (3G8, BD), anti-CD20 BUV737 (2H7, BD), anti-IgD BV605 (IA6-2, BD), and a fixable viability dye (Tonbo Biosciences, cat#13-0870-T500). B cells were individually sorted into flatbottom 96-well plates containing feeder cells that had been seeded at a density of 28,600 cells/well one day prior in 100 µL of IMDM media (Gibco, cat#31980030) containing 10% fetal calf serum, 100 U/mL penicillin plus 100 µg/mL streptomycin, and 2.5 µg/mL amphotericin. These 3T3 feeder cells produce CD40L, IL2, and IL21 to stimulate antibody secretion into culture supernatants [19 (link)]. B cells sorted onto feeder cells were cultured at 37 °C for 13 days.
Anti cd19 buv395 sj25c1
Manufactured by BD
Anti-CD19 BUV395 (SJ25C1) is a fluorochrome-conjugated monoclonal antibody that binds to the CD19 antigen, a transmembrane glycoprotein expressed on the surface of B cells. This product is intended for use in flow cytometry applications to identify and characterize CD19-positive cells.
Automatically generated - may contain errors
Lab products found in correlation
Facsaria, by BD (2 mentions)
Imdm media, by Thermo Fisher Scientific (2 mentions)
Anti igm fitc g20 127, by BD (2 mentions)
Anti cd3 bv711 ucht1, by BD (2 mentions)
Anti cd20 buv737 2h7, by BD (2 mentions)
Anti igd bv605 ia6 2, by BD (2 mentions)
Anti cd14 bv711 m0p 9, by BD (2 mentions)
Anti cd16 bv711 3g8, by BD (2 mentions)
2 protocols using anti cd19 buv395 sj25c1
1
B Cell Stimulation and Antibody Production
2
Isolation and Culture of B Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were incubated in 50 μL of FACS buffer containing a cocktail of antibodies for 30 min on ice prior to washing and analysis on a FACS Aria (BD). Antibodies included anti-IgM FITC (G20–127, BD, cat#555782, 1:80 dilution), anti-CD19 BUV395 (SJ25C1, BD, cat#563551, 1:20 dilution), anti-CD3 BV711 (UCHT1, BD, cat#563725, 1:50 dilution), anti-CD14 BV711 (M0P-9, BD, cat#563372, 1:50 dilution), anti-CD16 BV711 (3G8, BD, cat#563127, 1:50 dilution), anti-CD20 BUV737 (2H7, BD, cat#612849, 1:20 dilution), anti-IgD BV605 (IA6–2, BD, cat#563313, 1:50 dilution), anti-CD27 PE/Cy7 (LG.7F9, eBioscience, cat#25-0271-82, 1:160 dilution), and a fixable viability dye (Tonbo Biosciences, cat#13-0870-T500, 1:250 dilution). B cells were individually sorted into either 1) empty 96-well PCR plates and immediately frozen, or 2) flat bottom 96-well plates containing feeder cells that had been seeded at a density of 28,600 cells/well one day prior in 100 µL of IMDM media (Gibco, cat#31980030) containing 10% fetal calf serum, 100 U/mL penicillin plus 100 µg/mL streptomycin, and 2.5 µg/mL amphotericin. B cells sorted onto feeder cells were cultured at 37 °C for 13 days.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!