HUVEC were purchased from (Lonza) and were maintained using EGM2 medium (Lonza). HUVEC were used between passages 4 and 5. Jurkat T cells (E6.1, ATCC) and the corresponding CD47-deficient Jurkat mutant JinB8 were provided by Dr. Eric Brown, Genentech (Reinhold et al., 1999 (link)) and were maintained at 2.0 × 105 cells per ml in RPMI 1640 medium (Life Technologies) supplemented with 10% FBS, glutamine, penicillin, and streptomycin. Jurkat cells were maintained for a maximum of 4 weeks for experiments. Human anti-CD3 and anti-CD47-FITC (BD Biosciences), anti-CD69 (R&D systems), anti-actin, anti-tubulin, anti-VEGFR2, anti-VEGFR2 Y1175 (Cell Signaling), anti-CD47 (B6H12, Abcam) and azide-free functional grade anti-CD47 (e-Biosciences) were purchased from the indicated vendors.
Anti cd47 b6h12
Manufactured by Abcam
Sourced in United Kingdom
Anti-CD47 (B6H12) is a mouse monoclonal antibody that recognizes the CD47 antigen. CD47 is a cell surface protein involved in the regulation of phagocytosis.
Automatically generated - may contain errors
Lab products found in correlation
Rpmi 1640 medium, by Thermo Fisher Scientific (2 mentions)
Human umbilical vein endothelial cells (huvec), by Lonza (2 mentions)
Anti tubulin, by Cell Signaling Technology (2 mentions)
Anti actin, by Cell Signaling Technology (2 mentions)
Anti vegfr2, by Cell Signaling Technology (2 mentions)
Human anti cd3 and anti cd47 fitc, by BD (2 mentions)
Anti cd69, by R&D Systems (2 mentions)
Anti vegfr2 y1175, by Cell Signaling Technology (2 mentions)
Anti cd47, by Thermo Fisher Scientific (2 mentions)
Egm 2 medium, by Lonza (2 mentions)
Pkh67 green fluorescent cell linker kit, by Merck Group (1 mentions)
M csf, by Thermo Fisher Scientific (1 mentions)
3 protocols using anti cd47 b6h12
1
HUVEC and Jurkat Cell Maintenance
2
HUVEC and Jurkat Cell Maintenance
3
In Vitro Phagocytosis of NPC Cells by Macrophages
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Monocytes (human peripheral blood-derived mononuclear cells) were obtained from healthy donors after informed consent (LDEBIO, Guangzhou, China). Monocyte-derived macrophage cultures were established in RPMI 1640 supplemented with 10% FBS and 50 ng/ml M-CSF (eBioscience, USA) for 7 days.
For in vitro phagocytosis analysis, 1 × 104 monocyte-derived macrophages were plated in each well of 48-well tissue culture plates and labeled with a PKH67 Green Fluorescent Cell Linker Kit (Sigma-Aldrich, St. Louis, MO, USA) according to the manufacturer's instructions. Macrophages were incubated in serum-free medium for 2 h followed by an addition of 1 × 104 PKH26-labeled NPC cells. Anti-CD47 (B6H12, 10 mg/ml, Abcam, Cambridge, UK) and IgG1 control antibodies, respectively, were then added, and cells were incubated for 3 h at 37°C. After washing in PBS, the cells were fixed with 0.4% paraformaldehyde. The phagocytic index was assessed under a fluorescence microscope (lx71, Olympus, Japan) as the number of red phagocytosed NPC cells per 100 green macrophages in randomly selected microscopic fields.
For in vitro phagocytosis analysis, 1 × 104 monocyte-derived macrophages were plated in each well of 48-well tissue culture plates and labeled with a PKH67 Green Fluorescent Cell Linker Kit (Sigma-Aldrich, St. Louis, MO, USA) according to the manufacturer's instructions. Macrophages were incubated in serum-free medium for 2 h followed by an addition of 1 × 104 PKH26-labeled NPC cells. Anti-CD47 (B6H12, 10 mg/ml, Abcam, Cambridge, UK) and IgG1 control antibodies, respectively, were then added, and cells were incubated for 3 h at 37°C. After washing in PBS, the cells were fixed with 0.4% paraformaldehyde. The phagocytic index was assessed under a fluorescence microscope (lx71, Olympus, Japan) as the number of red phagocytosed NPC cells per 100 green macrophages in randomly selected microscopic fields.
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!