Mab307

Manufactured by R&D Systems

MAb307 is a monoclonal antibody product developed by R&D Systems for use in research applications. It is a purified immunoglobulin G (IgG) antibody. The core function of MAb307 is to serve as a tool for the detection and study of its target antigen, although specific details about the target are not provided.

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Lab products found in correlation

7 aminoactinomycin d 7 aad, by Thermo Fisher Scientific (1 mentions) Rhuepo, by R&D Systems (1 mentions) Interleukin 6 (il 6), by R&D Systems (1 mentions) Interleukin 3 (il 3), by R&D Systems (1 mentions) Amicon ultra 0.5 ml centrifugal filter, by Merck Group (1 mentions) Anti tgf β1, by Cell Signaling Technology (1 mentions) Anti tsg101, by Thermo Fisher Scientific (1 mentions) Af1180, by R&D Systems (1 mentions)

2 protocols using mab307

Erythroid Progenitor Cells Differentiation

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Human CD34+ progenitor cells (AllCells Inc., Emeryville, CA) were isolated from bone marrow using CD34 immunomagnetic purification (Miltenyi Biotec, Auburn, CA), per the manufacturer’s protocol. Differentiation of EPCs was induced with rHuEpo (0.1 U/mL), IL-3, IL-6, and stem cell factor (SCF) (R&D Systems, Minneapolis, MN). The use of CD36+/CD34- expression as markers for erythroid lineage development has been previously described [19 (link), 30 (link)]. EpoR function was analyzed by exposing the culture at various time points to a range of rHuEpo concentrations from 0 U/mL (rHuEpo formulation buffer “vehicle” control: 100 mM NaCl, 20 mM NaCitrate, 0.25% human serum albumin [HAS], pH 6.9) to 300 U/mL for 5 or 30 minutes, which includes the physiological range of endogenous Epo (5–30 mU/mL) and exceeds the pharmacological levels observed in patients treated with ESAs (reported to be a mean of 1 U/mL in plasma) [31 ]. Thus, the upper range of the titration was at least 300-fold higher than the theoretical exposure of tumor cells in patients administered ESAs. EpoR cell-surface expression was determined by flow cytometry using the EpoR specific antibody MAb307 (R&D Systems) incorporating 7-amino-actinomycin-D (7-AAD; Invitrogen) staining to select live cells with intact plasma membranes.

Patterson S.D., Rossi J.M., Paweletz K.L., Fitzpatrick V.D., Begley C.G., Busse L., Elliott S, & McCaffery I. (2015). Functional EpoR Pathway Utilization Is Not Detected in Primary Tumor Cells Isolated from Human Breast, Non-Small Cell Lung, Colorectal, and Ovarian Tumor Tissues. PLoS ONE, 10(3), e0122149.

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Exosomal Marker Profiling in Leukemia

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In preparation for Western blots, exosome fraction #4 was concentrated by centrifugation on a 100 K Amicon Ultra 0.5 mL centrifugal filter (EMD Millipore, Billerica, MA, USA) at 5000 × g. Western blots were performed as previously described [12 (link)]. PVD membranes were incubated overnight at 4 °C with various antibodies (Abs) as indicated below. Exosomes (10 μg protein) were loaded in each lane and tested for the presence of exosome markers, including markers related to hematopoiesis, leukemia blasts markers, and Leukemia Associated Antigens (LAAs) as previously described [12 (link)]. The following Abs were used: anti-TGF-β1 (Cell Signaling, #3711, 1:1000); anti-TNF-a (Cell Signaling, 3707S, 1:1000); anti-TSG101 (Thermo Fisher, PA5–31260, 1:500); anti-EPOR (R&D, MAB307, 1:500); anti-CD26 (R&D, AF1180, 1:500); anti-CLL-1 (R&D, AF2946, 1:2000); anti-CD34 (Santa Cruz, sc-7045, 1:500); and anti-CD33 (Thermo Fisher, WM53, 1:500). Band intensities on exposed films were quantified using Image J software (NIH, USA). The examined band intensity was normalized to the intensity of TSG101 used as a marker of the exosome endocytic origin. The integrated pixel value was determined for each protein band by multiplying image intensity and band area after subtracting the mean background value.

Namburi S., Broxmeyer H.E., Hong C.S., Whiteside T.L, & Boyiadzis M. (2020). DPP4+ exosomes in AML patients’ plasma suppress proliferation of hematopoietic progenitor cells. Leukemia, 35(7), 1925-1932.

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