Journal: Methods (San Diego, Calif.)

Article Title: Isolation and analysis of rare cells in the blood of cancer patients using a negative depletion methodology

doi: 10.1016/j.ymeth.2013.09.006

Figure Lengend Snippet: Flow chart of the CD45-based negative depletion and analysis pathway.

Article Snippet: The immunomagnetic labeling protocol involves two reagents: a CD45 tetrameric antibody complex (TAC) (Stem Cell Technologies) and dextran-coated magnetic nanoparticles (Stem Cell Technologies).

Techniques:

Journal: Methods (San Diego, Calif.)

Article Title: Isolation and analysis of rare cells in the blood of cancer patients using a negative depletion methodology

doi: 10.1016/j.ymeth.2013.09.006

Figure Lengend Snippet: Table of primary antibodies.

Article Snippet: The immunomagnetic labeling protocol involves two reagents: a CD45 tetrameric antibody complex (TAC) (Stem Cell Technologies) and dextran-coated magnetic nanoparticles (Stem Cell Technologies).

Techniques:

Journal: Methods (San Diego, Calif.)

Article Title: Isolation and analysis of rare cells in the blood of cancer patients using a negative depletion methodology

doi: 10.1016/j.ymeth.2013.09.006

Figure Lengend Snippet: List of CD45 antibody clones, fluoroprobes, and manufacturers tested.

Article Snippet: The immunomagnetic labeling protocol involves two reagents: a CD45 tetrameric antibody complex (TAC) (Stem Cell Technologies) and dextran-coated magnetic nanoparticles (Stem Cell Technologies).

Techniques: Clone Assay

Journal: Methods (San Diego, Calif.)

Article Title: Isolation and analysis of rare cells in the blood of cancer patients using a negative depletion methodology

doi: 10.1016/j.ymeth.2013.09.006

Figure Lengend Snippet: (A) Histograms of various anti-CD45 antibody clones, based on FCM analysis of peripheral blood leukocytes, both before (blue) and after (red) immunomagnetic labeling with anti-CD45-TAC/magnetic nanoparticles. Unstained controls are shown in orange. (B) Bar graph comparing the percentage of cells positive for each clone of anti-CD45 antibody, based on FCM analysis, both before and after immunomagnetic labeling.

Article Snippet: The immunomagnetic labeling protocol involves two reagents: a CD45 tetrameric antibody complex (TAC) (Stem Cell Technologies) and dextran-coated magnetic nanoparticles (Stem Cell Technologies).

Techniques: Clone Assay, Labeling

Journal: Methods (San Diego, Calif.)

Article Title: Isolation and analysis of rare cells in the blood of cancer patients using a negative depletion methodology

doi: 10.1016/j.ymeth.2013.09.006

Figure Lengend Snippet: Four-color images of a cytospin stained for DAPI, CK, CD45, and EpCAM of (a–e) leukocytes, (f–j) breast cancer cell line MCF7, and (k–o) an enriched peripheral blood sample from a metastatic breast cancer patient. The first column is a combined image of the four colors and the remaining columns (left to right) are DAPI, anti-CK-AF488, anti-CD45-AF594, and anti-EpCAM-AF647, respectively. For the patient sample, Cell #1 is CK+CD45+EpCAM− and Cell #2 is CK−CD45+EpCAM+.

Article Snippet: The immunomagnetic labeling protocol involves two reagents: a CD45 tetrameric antibody complex (TAC) (Stem Cell Technologies) and dextran-coated magnetic nanoparticles (Stem Cell Technologies).

Techniques: Staining

Journal: Methods (San Diego, Calif.)

Article Title: Isolation and analysis of rare cells in the blood of cancer patients using a negative depletion methodology

doi: 10.1016/j.ymeth.2013.09.006

Figure Lengend Snippet: (A) Four-color images of a cytospin stained for DAPI, CK, CD45, and VIM of (a–e) leukocytes, and (f–j) an enriched peripheral blood sample from a cancer patient. The columns (left to right) are DAPI, anti-CK-AF488, anti-CD45-AF594, anti-VIM-AF647, and a combined image of the four colors, respectively. (B) Four-color image of a cytospin stained for DAPI, CK, Vim, and N-CAD of an enriched peripheral blood sample from a cancer patient. (a) DAPI; (b) anti-CK-AF488; (c) anti-Vim-AF555; (d) anti-N-CAD-AF633 and (e) combined image of the four colors. The cells in the yellow boxes are CK+Vim+N-CAD+. Reproduced with permission from Balasubramanian et al. [15].

Article Snippet: The immunomagnetic labeling protocol involves two reagents: a CD45 tetrameric antibody complex (TAC) (Stem Cell Technologies) and dextran-coated magnetic nanoparticles (Stem Cell Technologies).

Techniques: Staining

Journal: Methods (San Diego, Calif.)

Article Title: Isolation and analysis of rare cells in the blood of cancer patients using a negative depletion methodology

doi: 10.1016/j.ymeth.2013.09.006

Figure Lengend Snippet: Four-color image of a cytospin stained for DAPI, CK, CD45, and HER2 of an enriched peripheral blood sample from a metastatic breast cancer patient. (a) DAPI; (b) anti-CK-AF488; (c) anti-CD45-AF594; (d) anti-HER2-AF647 and (e) combined image of the four colors. The cell in this figure is CK+CD45−HER2+, a HER2-expressing CTC.

Article Snippet: The immunomagnetic labeling protocol involves two reagents: a CD45 tetrameric antibody complex (TAC) (Stem Cell Technologies) and dextran-coated magnetic nanoparticles (Stem Cell Technologies).

Techniques: Staining, Expressing

Journal:

Article Title: Transferrin receptor 2 protein is not expressed in normal erythroid cells

doi: 10.1042/BJ20040230

Figure Lengend Snippet: Representative results on freshly purified human CD34+ progenitor cells induced to uni-lineage erythroid differentiation (E) and analysed on different days of culture (from day 0 to day 13). Aliquots of reverse-transcribed RNA from 5×104 cells were collected at the indicated days and amplified by 38 PCR cycles based on glyceraldehyde-3-phosphate dehydrogenase (GAPDH) normalization. K562 cells were used as a positive control.

Article Snippet: Human CD34 + progenitor cells were purified from peripheral blood by positive selection using the midi-MACS immunomagnetic separation system (Miltenyi Biotec, Bergisch Gladbach, Germany) according to the manufacturer's instructions.

Techniques: Purification, Reverse Transcription, Amplification, Positive Control

Journal:

Article Title: Transferrin receptor 2 protein is not expressed in normal erythroid cells

doi: 10.1042/BJ20040230

Figure Lengend Snippet: K562 cells, HepG2 cells, purified CD34+ cells, immature and mature erythroblasts were labelled with either G/14C2 or G/14E8 anti-TFR2 mAbs using an indirect immunofluorescence technique, and analysed for fluorescence emission using a flow cytometer. Immature and mature erythroblasts were derived from erythroid uni-lineage cultures at days 7 and 14 respectively. Filled-in histograms indicate the fluorescence observed in cells incubated with irrelevant mouse Igs (negative control); histograms without shading show the fluorescence observed in cells incubated with anti-TFR2 mAbs.

Article Snippet: Human CD34 + progenitor cells were purified from peripheral blood by positive selection using the midi-MACS immunomagnetic separation system (Miltenyi Biotec, Bergisch Gladbach, Germany) according to the manufacturer's instructions.

Techniques: Purification, Immunofluorescence, Fluorescence, Flow Cytometry, Derivative Assay, Incubation, Negative Control