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Cd9 pe

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CD9-PE is a fluorescent-labeled antibody that binds to the CD9 antigen, a member of the tetraspanin family of cell surface proteins. It is commonly used in flow cytometry applications to identify and characterize cells expressing the CD9 antigen.

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Lab products found in correlation

Permeabilization buffer, by Thermo Fisher Scientific (2 mentions) Paraformaldehyde (pfa), by Electron Microscopy Sciences (2 mentions) Cisplatin, by Merck Group (2 mentions) Trustain fcx fc receptor block, by BioLegend (2 mentions) Anti pe 165ho, by Standard BioTools (2 mentions) Withcd9 156gd, by Standard BioTools (2 mentions) Cd9 bv421, by BD (2 mentions) Live dead fixable near ir stain, by Thermo Fisher Scientific (2 mentions) Efluor 780, by Thermo Fisher Scientific (1 mentions) Foxp3 transcription factor staining buffer set, by Thermo Fisher Scientific (1 mentions) Cd4 fitc, by BD (1 mentions) Cd3 pe, by BD (1 mentions) Ifn γ fitc, by BD (1 mentions) Golgistop, by BD (1 mentions) Brilliant stain buffer, by BD (1 mentions) 7 aad, by BioLegend (1 mentions) Cd4 bv786, by BD (1 mentions) Cd45ra bv711, by BD (1 mentions) Cd44 pe, by BD (1 mentions) Cd86 pe, by BD (1 mentions)

Spelling variants of this product

PE-labeled anti-CD9 (2 citations) PE mouse anti-human CD9 (2 citations)

7 protocols using cd9 pe

1

Multiparametric Analysis of CD9 Expression

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NK-92 and OVCAR4 cells were stained with cisplatin (Sigma Aldrich),
fixed with 1.6% paraformaldehyde (Electron Microscopy Sciences), washed,
incubated with TruStain FcX Fc receptor block (BioLegend) (as above) and
then incubated with CD9-PE (Becton Dickinson) for 45min at room temperature.
Cells were washed and stained with anti-PE-165Ho (Fluidigm) for 30min at
room temperature. Following secondary antibody staining, cells were
permeabilized with 1x Permeabilization Buffer (eBioscience, Thermo Fisher
Scientific), on ice for 10min. Cells were subsequently stained with
CD9–156Gd (Fluidigm) (to detect intracellular CD9) for 1h at room
temperature. Cells were processed and introduced into the CyTOF 2 as
described above.
Gonzalez V.D., Huang Y.W., Delgado-Gonzalez A., Chen S.Y., Donoso K., Sachs K., Gentles A.J., Allard G.M., Kolahi K.S., Howitt B.E., Porpiglia E, & Fantl W.J. (2021). High-grade serous ovarian tumor cells modulate NK cell function to create an immune-tolerant microenvironment. Cell reports, 36(9), 109632.
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2

Multiparametric Analysis of CD9 Expression

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NK-92 and OVCAR4 cells were stained with cisplatin (Sigma Aldrich),
fixed with 1.6% paraformaldehyde (Electron Microscopy Sciences), washed,
incubated with TruStain FcX Fc receptor block (BioLegend) (as above) and
then incubated with CD9-PE (Becton Dickinson) for 45min at room temperature.
Cells were washed and stained with anti-PE-165Ho (Fluidigm) for 30min at
room temperature. Following secondary antibody staining, cells were
permeabilized with 1x Permeabilization Buffer (eBioscience, Thermo Fisher
Scientific), on ice for 10min. Cells were subsequently stained with
CD9–156Gd (Fluidigm) (to detect intracellular CD9) for 1h at room
temperature. Cells were processed and introduced into the CyTOF 2 as
described above.
Gonzalez V.D., Huang Y.W., Delgado-Gonzalez A., Chen S.Y., Donoso K., Sachs K., Gentles A.J., Allard G.M., Kolahi K.S., Howitt B.E., Porpiglia E, & Fantl W.J. (2021). High-grade serous ovarian tumor cells modulate NK cell function to create an immune-tolerant microenvironment. Cell reports, 36(9), 109632.
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3

Multiparametric Flow Cytometry Immunophenotyping

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For surface staining, antibodies were diluted in brilliant stain buffer (BD Biosciences, P/N 563794) and cells were stained with this mixture. Washing steps were performed with a standard flow cytometry buffer (1% FBS, 0.02% NaN3, PBS). For intracellular staining, the Foxp3/Transcription Factor Staining Buffer Set (Invitrogen, P/N 00-5523-00) was used according to the manufacturer’s instructions. Cells were pretreated with GolgiStop for cytokine staining (BD, P/N 554724). Viability was assessed using either 7-AAD (BioLegend, P/N 420403) or fixable viability dye eFluor780 (Invitrogen, P/N 65-0865-14). All antibodies used were from BD Biosciences: CD3-PE (P/N 566683), CD4-FITC (P/N 300538), CD4-BV786 (P/N 563877), IFN-γ-FITC (P/N 554551), Isotype control-FITC (P/N 554679), CD9-PE (P/N 555372), CD30-BV421 (P/N 562876), CD45RA-BV711 (P/N 612847), CD45RO-BV421 (P/N 562641), CD52-AF647 (P/N 563610), CD96-BV711 (P/N 563174).
Ford S.L., Buus T.B., Nastasi C., Geisler C., Bonefeld C.M., Ødum N, & Woetmann A. (2023). In vitro differentiated human CD4+ T cells produce hepatocyte growth factor. Frontiers in Immunology, 14, 1210836.
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4

Multicolor Flow Cytometry Protocol

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Antibodies against CD9 BV421, CD9 PE, from BD and CD45 APC from
BioLegend were used to detect CD9 and CD45. The same antibody clones were
used for CyTOF (Tables
S3 and S7). Near-IR fixable LIVE/DEAD stain from Thermo Fisher
Scientific was used to distinguish dead cells.
Gonzalez V.D., Huang Y.W., Delgado-Gonzalez A., Chen S.Y., Donoso K., Sachs K., Gentles A.J., Allard G.M., Kolahi K.S., Howitt B.E., Porpiglia E, & Fantl W.J. (2021). High-grade serous ovarian tumor cells modulate NK cell function to create an immune-tolerant microenvironment. Cell reports, 36(9), 109632.
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5

Multicolor Flow Cytometry Protocol

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Antibodies against CD9 BV421, CD9 PE, from BD and CD45 APC from
BioLegend were used to detect CD9 and CD45. The same antibody clones were
used for CyTOF (Tables
S3 and S7). Near-IR fixable LIVE/DEAD stain from Thermo Fisher
Scientific was used to distinguish dead cells.
Gonzalez V.D., Huang Y.W., Delgado-Gonzalez A., Chen S.Y., Donoso K., Sachs K., Gentles A.J., Allard G.M., Kolahi K.S., Howitt B.E., Porpiglia E, & Fantl W.J. (2021). High-grade serous ovarian tumor cells modulate NK cell function to create an immune-tolerant microenvironment. Cell reports, 36(9), 109632.
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6

Comprehensive Antibody Characterization for Cell Analysis

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Affinity-purified F(ab′)2 fragments of polyclonal goat anti-mouse IgM (anti-Ig) and polyclonal goat anti-rabbit Ab-APC were obtained from Jackson ImmunoResearch Laboratories. Anti-mouse mAbs against CD19-APC, B220-PE, B220-FITC, CD5-PE-Cy7, CD43-PE, PD-L2-PE, and PD-L2-APC, CD86-PE, CD44-PE, CD9-PE, CD80-PE, Thy-1-PE, CD45.1-FITC, phospho-p38, and phospho-ERK-APC were obtained from BD pharmingen. Polyclonal anti-pERK, anti-NFATc1, anti-RasGRP3, and anti-phospho-JNK antibodies were obtained from Cell Signaling Technology. Anti-RasGRP1 antibody was obtained from Santa Cruz Biotechnology. Phorbol ester myristate (PMA), RIPA buffer, phenylmethylsulfonyl fluoride (PMSF), and protein inhibitor cocktails were obtained from Sigma Aldrich.
Guo B, & Rothstein T.L. (2016). RasGRP1 Is an Essential Signaling Molecule For Development of B1a Cells With Autoantigen Receptors. Journal of immunology (Baltimore, Md. : 1950), 196(6), 2583-2590.
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7

Tear Fluid Exosome Characterization by Flow Cytometry

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Flow cytometry was performed on a pooled tear fluid sample. Aliquots of 20 μL Dynabeads® (Exosome-Human CD9 Flow Detection Reagent, Invitrogen, Thermo Fisher Scientific, Oslo, Norway, Cat no 10620D) were added to 100 μL of the tear fluid sample and to each of three controls (negative control: assay buffer; positive control: SW−480 cell line EVs; ISO-control: tear fluid). Overnight incubation was performed in a HulaMixer Sample Mixer at 2–8 °C. Following wash steps, 150 μL of assay buffer was used to redistribute the beads, and 100 μL from each sample was then combined with 25 μL of PE-conjugated detection antibodies (CD9-PE, Cat no 555372, BD Biosciences, Oslo, Norway) or 25 μL of isotype-matched control (IgG1-PE, Cat no 559320, BD Biosciences, Oslo, Norway). An orbital shaker (IKA-WERKE GMBH & CO, Staufen, Germany) was then used for 45 min at 1000 rpm. Following two wash steps, a final 100 μL of assay buffer was added to each tube. All samples and controls were then placed on ice, ready for flow cytometry analysis. A BD Accuri™ C6 Cytometer (BD Biosciences, Oslo, Norway) was used for flow cytometry EV detection.
Cross T., Øvstebø R., Brusletto B.S., Trøseid A.M., Olstad O.K., Aspelin T., Jackson C.J., Chen X., Utheim T.P, & Haug K.B. (2023). RNA Profiles of Tear Fluid Extracellular Vesicles in Patients with Dry Eye-Related Symptoms. International Journal of Molecular Sciences, 24(20), 15390.
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