Mouse Anti-CD47 (B6H12) Recombinant Antibody

Expression Host：Nicotiana benthamiana plants

Clonality：Monoclonal, recombinant

Species and Isotype： Mouse IgG2a

Description：This product is a full-length Mouse IgG2a recombinant antibody specific to Human CD47 (IAP) protein. It was produced 

                      in Nicotiana benthamiana plants via Agrobacterium tumefaciens-mediated infiltration.

Verified Applications：Western blot, ELISA, Immunocytochemistry, Flow cytometry

Dilution Range：Western blot (1: 1 000 – 1: 5 500); ELISA (1: 3 000 – 1: 256 000); ICC (1:100-1:300); FC (1:300-1:500)

Tested Species Reactivity ： Human

Concentration ：1 mg/ml

Form ： Liquid

Storage：Short-term (up to one week): 2 – 8 °C

Long term: Aliquot and store at – 20 °C

Store immediately. Aliquot and avoid multiple freeze-thaw cycles.

Storage Buffer： 0.1   M Phosphate Buffered Saline, pH 7.4. Preservative: None

Purification Notes： This product was purified using Protein A- affinity chromatography.

Purity：≥ 95% as determined by SDS-PAGE

General Notes：For Research Use only, unless otherwise indicated.

Image：

Figure 1. ELISA Dose Response curve using Mouse Anti-CD47 (B6H12) antibody from 0.3 – 0.00014 ng/µl to detect 25 ng Human CD47 

protein. Experiments were performed in triplicate, with error bars representing standard deviation (SD).

Figure 2. Western blot of decreasing amounts of Human CD47 protein (~35-44 kDa smear) detected with Mouse Anti-CD47 (B6H12) antibody 

at 1: 5 000 and an HRP conjugated anti-mouse secondary antibody at 1: 10 000.

Figure 3. Immunocytochemistry: HeLa cells were plated at 200 000 cells/ well in 6-well plates on coverslips and allowed to adhere. Following

fixing and blocking, they were incubated with 1:200 dilution of Mouse Anti-CD47 (B6H12) primary antibody, and 1:300 dilution of a commercial 

Anti-Mouse Alexa Fluor® 488 conjugated commercial secondary antibody. Images were taken on a Zeiss LSM780 with ELYRA PS1 platform

confocal microscope (60X) at the Stellenbosch University CAF unit. Thank you to Prof Georgia Schafer (ICGEB) for kindly donating the HeLa 

cells and Mrs Lize Engelbrecht for her outstanding assistance.

Figure 4. Flow cytometry: Peripheral Blood Mononuclear cells (1.3 million cells) were fixed with 4% Paraformaldehyde and washed 2x in PBS

 buffer. Cells were stained with 1: 400 of Mouse Anti-CD47 Recombinant Antibody, followed by a commercial secondary Anti-Mouse 

conjugated to Alexa Fluor® 488 at 1: 400 dilution. Approximately 30 000 events were acquired on a BD FACSDiva 8.0.2 Flow cytometer. A)

 Dot-plot and B) Histogram of the acquired 30 000 events.