Immune Cell Subset Characterization

Excised tissues were prepared as previously described (41 (link)). Cell suspensions were blocked by the addition of Gamunex (Baxter, Deerfield, IL) for 30 minutes at RT, then washed in flow buffer (PBS + 2%FBS) and incubated at RT for 1 hour with the following antibodies (at a 1:10 final concentration): PerCP/Cy5.5-CD3, APC-CD34, PE-CD45, and/or APC-CD151 (BioLegend). Flow analysis was conducted on a Beckman Coulter Cyan. Cell sorting was performed on the Beckman Coulter Moflo XDP 70. Data analysis was performed using Summit, V5.1 (Beckman Coulter, Brea, CA).

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Morton J.J., Bird G., Keysar S.B., Astling D.P., Lyons T.R., Anderson R.T., Glogowska M.J., Estes P., Eagles J.R., Le P.N., Gan G., McGettigan B., Fernandez P., Padilla-Just N., Varella-Garcia M., Song J.I., Bowles D.W., Schedin P., Tan A.C., Roop D.R., Wang X.J., Refaeli Y, & Jimeno A. (2015). XactMice: humanizing mouse bone marrow enables microenvironment reconstitution in a patient-derived xenograft model of head and neck cancer. Oncogene, 35(3), 290-300.

Publication 2015

Gamunex PerCP/Cy5.5-CD3 APC-CD34 PE-CD45 APC-CD151 Moflo XDP 70 Summit, V5.1

Antibodies Buffer Cd151 Cell Cy5 5 Gamunex Tissues

Corresponding Organization :

Other organizations : University of Colorado Denver

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Variable analysis

independent variables

Cell type (e.g., CD3+, CD34+, CD45+, CD151+)

dependent variables

Cell population characteristics (e.g., cell counts, cell surface marker expression)

control variables

Gamunex blocking treatment (30 minutes at room temperature)
Antibody concentration (1:10 final concentration)
Incubation time (1 hour at room temperature)
Flow analysis platform (Beckman Coulter Cyan)
Cell sorting platform (Beckman Coulter Moflo XDP 70)
Data analysis software (Summit, V5.1)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

Annotations

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