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Pe labeled annexin 5

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PE labeled-Annexin V is a fluorescent protein-conjugated reagent used to detect and quantify apoptotic cells. Annexin V is a calcium-dependent phospholipid-binding protein that has a high affinity for phosphatidylserine, which is exposed on the surface of apoptotic cells. PE, or phycoerythrin, is a fluorescent dye used to label the Annexin V protein, allowing for the detection and analysis of apoptotic cells using flow cytometry or fluorescence microscopy.

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Lab products found in correlation

7 aad, by BD (7 mentions) Annexin 5 binding buffer, by BD (4 mentions) Brdu flow kit, by BD (2 mentions) Lsrfortessa flow cytometer, by BD (2 mentions) Facsaria 2, by BD (2 mentions) Fitc labeled goat anti mouse igg secondary antibody, by Abcam (1 mentions) Facscalibur flow cytometer, by BD (1 mentions) Mouse anti cd133 pe antibody clone ac133, by Miltenyi Biotec (1 mentions) Aldefluor kit, by STEMCELL (1 mentions) Anti survivin, by Cell Signaling Technology (1 mentions) Anti xiap, by Cell Signaling Technology (1 mentions) 3 4 5 dimethylthiazol 2 yl 2 5 diphenyltetrazolium bromide mtt, by Merck Group (1 mentions) Kms12bm, by Japanese Collection of Research Bioresources Cell Bank (1 mentions) Kms 11, by Japanese Collection of Research Bioresources Cell Bank (1 mentions) Kms 26, by Japanese Collection of Research Bioresources Cell Bank (1 mentions) Kms 12 pe, by Japanese Collection of Research Bioresources Cell Bank (1 mentions) Rpmi 8226, by Japanese Collection of Research Bioresources Cell Bank (1 mentions) Propidium iodide, by Merck Group (1 mentions) Facscalibur, by BD (1 mentions)

Close spelling variants of this product

7AAD and PE-labeled Annexin V PE-labeled Annexin V and 7-AAD Annexin V-PE Annexin V

11 protocols using pe labeled annexin 5

1

Quantification of HMGB1 and Annexin-V in Tobacco Smoke

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Monoclonal antibodies (mAb) against human HMGB1 (R&D Systems), phycoerythrin (PE)-labeled annexin-V (BD Pharmingen), and CRM1 inhibitor III (Calbiochem) were purchased. Tobacco smoke extract (TSE, 100%) was prepared by extracting mainstream smoke from lighted research cigarettes through phenol red-free RPMI-1640 medium containing 0.2% BSA (RPMI/BSA), then filtered (0.22-mm) and standardized according to its absorbance at 320 nm as described (Li et al., 2010 (link), 2013 (link)).
CHEN Y., LI G., LIU Y., WERTH V.P., WILLIAMS K.J, & LIU M.L. (2016). Translocation of Endogenous Danger Signal HMGB1 From Nucleus to Membrane Microvesicles in Macrophages. Journal of cellular physiology, 231(11), 2319-2326.
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2

Flow Cytometry Immunophenotyping and Cell Assays

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Approximately 106 cells per sample were resuspended in PBS blocked using Fc blocker for 10 minutes on ice, followed by staining with the appropriate dilution of the antibodies or their respective isotype controls for 15 minutes on ice. Cells were washed and resuspended in PBS with propidium iodide (PI, 0.2 μg/mL) or DAPI (0.75 μg/mL) as a dead cell marker. The antibodies used for flow cytometry are listed in Supplementary Table 7. For competitive growth assays, percentage of GFP+ cells was monitored by flow cytometry. For Annexin V staining, Annexin V binding buffer (BD Bioscience) was used instead of PBS and 7AAD (BD Bioscience) instead of PI. PE labeled-Annexin V was purchased from BD Bioscience. For BrdU staining, the BrdU Flow Kit was purchased from BD Bioscience and performed according to the manufacturer’s protocol.
Chan L.N., Chen Z., Braas D., Lee J.W., Xiao G., Geng H., Cosgun K.N., Hurtz C., Shojaee S., Cazzaniga V., Schjerven H., Ernst T., Hochhaus A., Kornblau S.M., Konopleva M., Pufall M.A., Cazzaniga G., Liu G.J., Milne T.A., Koeffler H.P., Ross T.S., Sanchez-Garcia I., Borkhardt A., Yamamoto K.R., Dickins R.A., Graeber T.G, & Müschen M. (2017). Metabolic gatekeeper function of B-lymphoid transcription factors. Nature, 542(7642), 479-483.
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3

Flow Cytometry Analysis of Microvesicles

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Flow cytometry of MVs and cells was performed according to our published methods (Liu et al., 2007 (link); Li et al., 2010 (link), 2013 (link)). In brief, culture supernatants were fixed in filtered 1% paraformaldehyde (PFA), and supplemented with counting beads. The number of MVs in each sample was quantified using a FACSCalibur flow cytometer, with gating criteria based on particle size and surface exposure of PS, detected by PE-labeled annexin-V (BD Pharmingen) staining (Liu et al., 2007 (link); Li et al., 2010 (link), 2013 (link)). The portion of PS-positive MVs that were also HMGB1-positive was quantified by simultaneous staining with mouse anti-human HMGB1 antibody (R&D), followed by FITC-labeled goat anti-mouse IgG secondary antibody (Abcam). After treatment without or with TSE, adherent monolayers of macrophages were detached and fixed by suspension in 1% PFA for detection of cell-surface HMGB1 with flow cytometry (Li et al., 2013 (link)).
CHEN Y., LI G., LIU Y., WERTH V.P., WILLIAMS K.J, & LIU M.L. (2016). Translocation of Endogenous Danger Signal HMGB1 From Nucleus to Membrane Microvesicles in Macrophages. Journal of cellular physiology, 231(11), 2319-2326.
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4

Cell Surface Staining and Flow Cytometry

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Approximately 106 cells per sample were resuspended in PBS with DAPI (0.75 μg/mL) as a dead cell marker. For competitive growth assays, percentage of GFP+ or KO1+ cells was monitored by flow cytometry. For cell surface staining, PBS washed cells were blocked with Fc blocker for 10 min on ice and then stained with the indicated antibodies listed in Supplementary Table S11 or isotype control for 25 min on ice. Cells were then washed and resuspended in chilled PBS containing 0.75 μg/mL of DAPI to exclude dead cells. For Annexin V staining, Annexin V binding buffer (BD Bioscience) was used instead of PBS and 7AAD (BD Bioscience) instead of DAPI. PE labeled-Annexin V was purchased from BD Bioscience. Acquisition was performed on LSRFortessa flow cytometer (BD Biosciences). Fluorescence-based cell sorting was performed on FACSAria II (BD Biosciences). FACS data were analyzed with FlowJo software (FlowJo, LLC).
Chan L.N., Murakami M.A., Robinson M.E., Caeser R., Sadras T., Lee J., Cosgun K.N., Kume K., Khairnar V., Xiao G., Ahmed M.A., Aghania E., Deb G., Hurtz C., Shojaee S., Hong C., Pölönen P., Nix M.A., Chen Z., Chen C.W., Chen J., Vogt A., Heinäniemi M., Lohi O., Wiita A.P., Izraeli S., Geng H., Weinstock D.M, & Müschen M. (2020). Signaling input from divergent pathways subverts B-cell transformation. Nature, 583(7818), 845-851.
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5

Lung CICs Isolation and Characterization

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Lung CICs were dissociated into single-cell suspensions by Accumaxtm cell counting solution (EMD Millipore) and stained with mouse anti-CD133-PE antibody (clone AC133, Miltenyi Biotec) and mouse anti-CD44-FITC antibody (clone DB105, Miltenyi Biotec). The cells were sorted by flow cytometry. Apoptosis was measured by flow cytometry analysis of PE-labeled Annexin-V (BD Biosciences) and 7-AAD (BD Biosciences) co-staining, according to the manufacturer’s protocol. ALDH assay was performed by flow cytometry analysis with ALDEFLUOR™ Kit (STEMCELL), following the recommendation of the manufacturer.
Yun X., Zhang K., Wang J., Pangeni R.P., Yang L., Bonner M., Wu J., Wang J., Nardi I.K., Gao M, & Raz D.J. (2018). Targeting USP22 Suppresses Tumorigenicity and Enhances Cisplatin Sensitivity Through ALDH1A3 Downregulation in Cancer-initiating Cells from Lung Adenocarcinoma. Molecular cancer research : MCR, 16(7), 1161-1171.
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6

Quantification of Apoptosis by Flow Cytometry

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Apoptosis was quantified using 7AAD and PE-labeled Annexin V (BD Biosciences) following the manufacturer’s protocol and flow cytometry. Cells were collected 48 h after transfection, washed twice with cold PBS, and resuspended at 1 × 106 cells/mL and mixed with 100 μL of 1× buffer and 5 μL Annexin V- PE and 7AAD, incubated for 15 min in the dark, 400 μL 1× buffer was added, and the cells were subjected to cytometry flow within 1 h.
Chen S., Sun K.X., Liu B.L., Zong Z.H, & Zhao Y. (2016). MicroRNA-505 functions as a tumor suppressor in endometrial cancer by targeting TGF-α. Molecular Cancer, 15, 11.
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7

Flow Cytometry Immunophenotyping and Cell Assays

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Approximately 106 cells per sample were resuspended in PBS blocked using Fc blocker for 10 minutes on ice, followed by staining with the appropriate dilution of the antibodies or their respective isotype controls for 15 minutes on ice. Cells were washed and resuspended in PBS with propidium iodide (PI, 0.2 μg/mL) or DAPI (0.75 μg/mL) as a dead cell marker. The antibodies used for flow cytometry are listed in Supplementary Table 7. For competitive growth assays, percentage of GFP+ cells was monitored by flow cytometry. For Annexin V staining, Annexin V binding buffer (BD Bioscience) was used instead of PBS and 7AAD (BD Bioscience) instead of PI. PE labeled-Annexin V was purchased from BD Bioscience. For BrdU staining, the BrdU Flow Kit was purchased from BD Bioscience and performed according to the manufacturer’s protocol.
Chan L.N., Chen Z., Braas D., Lee J.W., Xiao G., Geng H., Cosgun K.N., Hurtz C., Shojaee S., Cazzaniga V., Schjerven H., Ernst T., Hochhaus A., Kornblau S.M., Konopleva M., Pufall M.A., Cazzaniga G., Liu G.J., Milne T.A., Koeffler H.P., Ross T.S., Sanchez-Garcia I., Borkhardt A., Yamamoto K.R., Dickins R.A., Graeber T.G, & Müschen M. (2017). Metabolic gatekeeper function of B-lymphoid transcription factors. Nature, 542(7642), 479-483.
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8

Cell Surface Staining and Flow Cytometry

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Approximately 106 cells per sample were resuspended in PBS with DAPI (0.75 μg/mL) as a dead cell marker. For competitive growth assays, percentage of GFP+ or KO1+ cells was monitored by flow cytometry. For cell surface staining, PBS washed cells were blocked with Fc blocker for 10 min on ice and then stained with the indicated antibodies listed in Supplementary Table S11 or isotype control for 25 min on ice. Cells were then washed and resuspended in chilled PBS containing 0.75 μg/mL of DAPI to exclude dead cells. For Annexin V staining, Annexin V binding buffer (BD Bioscience) was used instead of PBS and 7AAD (BD Bioscience) instead of DAPI. PE labeled-Annexin V was purchased from BD Bioscience. Acquisition was performed on LSRFortessa flow cytometer (BD Biosciences). Fluorescence-based cell sorting was performed on FACSAria II (BD Biosciences). FACS data were analyzed with FlowJo software (FlowJo, LLC).
Chan L.N., Murakami M.A., Robinson M.E., Caeser R., Sadras T., Lee J., Cosgun K.N., Kume K., Khairnar V., Xiao G., Ahmed M.A., Aghania E., Deb G., Hurtz C., Shojaee S., Hong C., Pölönen P., Nix M.A., Chen Z., Chen C.W., Chen J., Vogt A., Heinäniemi M., Lohi O., Wiita A.P., Izraeli S., Geng H., Weinstock D.M, & Müschen M. (2020). Signaling input from divergent pathways subverts B-cell transformation. Nature, 583(7818), 845-851.
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9

Apoptosis Quantification via Flow Cytometry

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The cells were collected at 1500 r for 5 min and washed with PBS. 5 μL of 7AAD and PE-labeled Annexin V (BD Biosciences) were used to label apoptotic cells for each sample. The cells apoptosis rate was determined with flow cytometry.
Zong Z.H., Du Y.P., Guan X., Chen S, & Zhao Y. (2019). CircWHSC1 promotes ovarian cancer progression by regulating MUC1 and hTERT through sponging miR-145 and miR-1182. Journal of Experimental & Clinical Cancer Research : CR, 38, 437.
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10

Apoptosis Detection via Flow Cytometry

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Flow cytometry was performed with 7-amino-actinomycin (7-AAD) and phycoerythrin (PE)-labeled annexin V (BD Pharmingen, USA) to detect phosphatidylserine externalization as an endpoint indicator of early apoptosis as the protocol recommends.
Yun W.J., Li J., Yin N.C., Zhang C.Y., Cui Z.G., Zhang L, & Zheng H.C. (2024). The facilitating effects of KRT80 on chemoresistance, lipogenesis, and invasion of esophageal cancer. Cancer Biology & Therapy, 25(1), 2302162.
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