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Pe conjugated annexin 5

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PE-conjugated Annexin V is a fluorescent-labeled protein that binds to phosphatidylserine, a phospholipid that is exposed on the surface of apoptotic cells. This product can be used for the detection and quantification of apoptotic cells in flow cytometry applications.

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Lab products found in correlation

Facsdiva software, by BD (3 mentions) Facsaria, by BD (3 mentions) Fluo 4 am, by Thermo Fisher Scientific (2 mentions) Annexin 5 binding buffer, by BD (2 mentions) Facscalibur, by BD (2 mentions) 7 aminoactinomycin d, by BD (2 mentions) Propidium iodide, by BD (2 mentions) 7 aad, by BD (2 mentions) Facscanto 2, by BD (2 mentions) Annexin 5 fluos staining kit, by Roche (2 mentions) Matrigel matrix, by BD (1 mentions) Ebm 2 basal medium, by Lonza (1 mentions) Egm 2 singlequots kit, by Lonza (1 mentions) Vectashield antifade mounting medium, by Vector Laboratories (1 mentions) Uranyl acetate, by Electron Microscopy Sciences (1 mentions) Normal goat serum (ngs), by Vector Laboratories (1 mentions) Bovine thrombin, by Merck Group (1 mentions) Sepharose cl 2b column, by Merck Group (1 mentions) Lead nitrate, by Avantor (1 mentions) Rhosin hydrochloride, by Bio-Techne (1 mentions)

Spelling variants of this product

Annexin V (870 citations) Annexin V-PE (283 citations) PE-conjugated Annexin V and 7-AAD (6 citations) PE-conjugated anti-Annexin V and 7-AAD (6 citations) PE-conjugated Annexin V labeling (3 citations) PE-conjugated Annexin V antibody (3 citations) PE)-conjugated anti-annexin V antibody (2 citations)

33 protocols using pe conjugated annexin 5

1

Platelet activation and angiogenesis

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Acid-soluble thrombin receptor PAR1 activating peptide (PAR1-AP) was from Calbiochem (Darmstadt, Germany). Water-soluble PAR1-AP and PAR4 activating peptide (PAR4-AP), prostaglandin I2 (PGI2), and cultured cell detach solution (0.01% trypsin/5 mM EDTA) were purchased from Sigma (St Louis, MO, USA). Platelets were identified by the fluorescein isothiocyanate (FITC)-conjugated CD42a MAb Beb 1 (Becton Dickinson; San Jose, CA, USA). Platelet P-selectin expression was determined by R-phycoerythrin (PE)-CD62P MAb, and platelet phosphotidylserine (PS) exposure was monitored by PE-conjugated annexin V (both from Becton Dickinson). Immunoassay kits monitoring platelet release of vascular endothelial growth factor (VEGF), platelet-derived growth factor (PDGF, BB type), platelet factor 4 (PF4), and thrombospondin-1 (TSP-1) were from R&D Systems Ltd (Abingdon, UK). Endothelial culture media (EBM-2 Basal Medium and the EGM-2 SingleQuots kit) and fetal bovine serum (FBS) were purchased from Lonza (Basel, Switzerland). Matrigel ™ Matrix was from Becton Dickinson.
Miao X., Zhang W., Huang Z, & Li N. (2016). Unaltered Angiogenesis-Regulating Activities of Platelets in Mild Type 2 Diabetes Mellitus despite a Marked Platelet Hyperreactivity. PLoS ONE, 11(9), e0162405.
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2

Platelet Activation Signaling Pathway Analysis

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Sepharose CL‐2B column, bovine thrombin, and Ca2+‐ionophore calcimycin were from Sigma‐Aldrich. CVX was the product of Pentapharm. Phycoerythrin (PE)‐labeled anti‐CD62 antibody (Ab), PE‐cyanine5 (PECy5)‐conjugated anti‐CD41a Ab, PE‐labeled mouse IgG1, fluorescein isothiocyanate (FITC)‐labeled mouse IgG2a, PE‐conjugated annexin V, and annexin V binding buffer were purchased from Becton Dickinson. Mouse anti‐human CD41a Ab, Alexa‐fluor 568 annexin V conjugate, and DyLight 405‐labeled goat anti‐mouse Ab were the products of Thermo Fisher Scientific. DyLight 488‐labeled horse anti‐rabbit Ab, Vectashield® antifade mounting medium, and normal goat serum were from Vector Laboratories. Goat anti‐rabbit IgG conjugated to 15 nm gold particles and goat anti‐mouse IgG conjugated to 10 nm gold particles were purchased from BBI Solutions. Uranyl acetate was from Electron Microscopy Sciences. Lead citrate was made from lead nitrate (VWR International Ltd.). Rabbit anti‐human FXIII‐A Ab and FITC‐labeled mouse anti‐human‐FXIII‐A Ab were produced in our laboratories.16, 22 The fluorescent calcium indicator Fluo‐4‐AM was obtained from Thermo Fisher Scientific. The RhoA inhibitor Rhosin hydrochloride was purchased from Tocris Bioscience and the transglutaminase inhibitor T101 from Zedira.
Somodi L., Beke Debreceni I., Kis G., Cozzolino M., Kappelmayer J., Antal M., Panyi G., Bárdos H., Mutch N.J, & Muszbek L. (2022). Activation mechanism dependent surface exposure of cellular factor XIII on activated platelets and platelet microparticles. Journal of Thrombosis and Haemostasis, 20(5), 1223-1235.
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3

Assessing T Cell Apoptosis Induction

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T cell blasts were left unstimulated or stimulated for 12 h with 0.01, 0.1, 1, and 10 µg ml−1 immobilized anti-CD3 (clone OKT3) or cross-linked anti-FAS antibody (clone Apo1.3) as previously described (Rigaud et al., 2006 (link)). Cells were then washed and stained for viability (viaprobe; BD Biosciences); surface expression of CD3, CD4, and CD8; and surface localization of phosphatidylserines using PE-conjugated Annexin-V (BD Biosciences) and analyzed by flow cytometry. Apoptotic cells corresponding to Annexin V+/viaprobe− cells.
Rodriguez R., Fournier B., Cordeiro D.J., Winter S., Izawa K., Martin E., Boutboul D., Lenoir C., Fraitag S., Kracker S., Watts T.H., Picard C., Bruneau J., Callebaut I., Fischer A., Neven B, & Latour S. (2019). Concomitant PIK3CD and TNFRSF9 deficiencies cause chronic active Epstein-Barr virus infection of T cells. The Journal of Experimental Medicine, 216(12), 2800-2818.
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4

Nutlin-3a Induced Apoptosis in SiHa and HEK Cells

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SiHa cells or HEK cells, cultured in 96-well plates at a concentration of 4.0 × 103/well or 8.0 × 103/well, were transduced or transfected following the above steps. Then they were treated with or without 40 µg ml−1 or 20 µg ml−1 of nutlin-3a for one to five days. Cellular viability assay was detected using water-soluble tetrazolium 1 (WST-1) assay at each time point according to the protocol. Each treatment was performed in quadruplicate, and the experiment was repeated three times.
At 72 hpt, the cells in 6-well plates were harvested and resuspended in 100 µl of 1 × Annexin-V binding buffer. The supernatant was incubated with 5 µl of phycoerythrin (PE)-conjugated Annexin V (BD Pharmingen, USA) and 5 µl of 7-amino-actinomycin (7-AAD, BD Pharmingen) for 15 min at room temperature in the dark, and then 400 µl of 1 × buffer was added to each tube. Flow cytometric analysis (BD FACSCalibur, BD Biosciences, USA) was performed within 1 h of staining. Each experiment was performed with at least three biological replicates.
Liu X., Zhang Y., Wang S., Liu G, & Ruan L. (2018). Loss of miR-143 and miR-145 in condyloma acuminatum promotes cellular proliferation and inhibits apoptosis by targeting NRAS. Royal Society Open Science, 5(8), 172376.
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5

Quantifying Cellular Viability, Apoptosis, and Clonogenicity

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Number of viable cells was enumerated by PrestoBlue assay (Lam et al, 2016). Apoptosis was performed using PE‐conjugated Annexin V (BD Bioscience) and 7‐aminoactinomycin D (7‐AAD) according to the manufacturer's instructions. Clonogenicity was measured by standard methylcellulose‐based system (MethoCult, Stem Cell Technologies). AML cells (100 cells for MOLM‐13, 200 cells for MV4‐11, and 500 cells for ML‐2) were seeded in triplicates in 35‐mm culture plates. Colonies were stained with Giemsa (Sigma) and enumerated after 10 days of culture.
He B., Yang N., Man C.H., Ng N.K., Cher C., Leung H., Kan L.L., Cheng B.Y., Lam S.S., Wang M.L., Zhang C., Kwok H., Cheng G., Sharma R., Ma A.C., So C.E., Kwong Y, & Leung A.Y. (2020). Follistatin is a novel therapeutic target and biomarker in FLT3/ITD acute myeloid leukemia. EMBO Molecular Medicine, 12(4), e10895.
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6

Cell Cycle and Apoptosis Analysis

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For cell cycle analysis, the lentiviral vector-transduced A549 and H1299 cells were labeled with propidium iodide (BD Biosciences, San Jose, CA, USA) and analyzed using flow cytometry. For apoptosis analysis, the cells were incubated with PE-conjugated annexin V and 7-AAD (BD Biosciences, San Jose, CA, USA), according to the manufacturer's guidelines, prior to flow cytometry.
Di X., Jin X., Ma H., Wang R., Cong S., Tian C., Liu J., Zhao M., Li R, & Wang K. (2019). The Oncogene IARS2 Promotes Non-small Cell Lung Cancer Tumorigenesis by Activating the AKT/MTOR Pathway. Frontiers in Oncology, 9, 393.
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7

Flow Cytometric Analysis of Apoptosis

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Cells were washed twice with PBS before incubation with cocktail antibodies consisting of FITC-conjugated anti-CD71, APC-conjugated anti-CD235a, PE-conjugated annexin V and 7-aminoactinomycin D (7-AAD) (all from BD Biosciences, USA) in binding buffer for 15 min at room temperature in the dark. Samples were analyzed by the FACS Canto flow cytometer (BD Biosciences, USA).
Klaihmon P., Lorthongpanich C., Kheolamai P., Saisaard W, & Issaragrisil S. (2024). Inhibition of LATS kinases reduces tumorigenicity and increases the sensitivity of human chronic myelogenous leukemia cells to imatinib. Scientific Reports, 14, 3993.
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8

Antibody Panel for Flow Cytometric Analysis

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The following antibodies were purchased from eBioscience (San Diego, CA) for flow cytometry: fluorescein isothiocyanate (FITC)-conjugated antibodies against CD11b (M1/70), CD4 (Gk1.50), and Rat IgG2b isotype control; phycoerythrin (PE)-conjugated antibodies against TLR4 (MTS510), RP105 (RP/14), CD80 (16-10A1), CD86 (GL1), MHC class I (AF6-88.5.5.3), MHC class II (M5/114.15.2), CD11b (M1/70), and isotype control; allophycocyanin (APC)-conjugated antibodies against CD8 (Ly-2), VSIG4 (NLA14), and isotype control; and purified anti-CD16/32 (2.4G2). Functional grade anti-TNF-α (MP6-XT22) was purchased from eBioscience for T-cell activation. 7-AAD was purchased from eBioscience and PE-conjugated Annexin V was purchase from BD Bioscience (Franklin Lakes, NJ) for apoptosis.
Cho H.Y., Yang Y.G., Jeon Y., Lee C.K., Choi I, & Lee S.W. (2021). VSIG4(+) peritoneal macrophages induce apoptosis of double-positive thymocyte via the secretion of TNF-α in a CLP-induced sepsis model resulting in thymic atrophy. Cell Death & Disease, 12(6), 526.
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9

Immunophenotyping and Apoptosis Analysis

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Fc receptors were blocked with anti-CD16+CD32 mAb (clone 2.4G2) before staining with the indicated mAbs (BD, eBioscience, or BioLegend). We assessed apoptosis by staining freshly isolated splenocytes and hepatic lymphocytes with PE-conjugated annexin V (BD) or using a probe for activated caspase (FLICA; Immunochemistry Technologies). Cell division was measured by injecting mice intravenously with 200 µg BrdU in PBS (BD) 2 h before tissue harvest. Samples were analyzed on an LSRII (BD) using FlowJo software (Tree Star).
Min-Oo G., Bezman N.A., Madera S., Sun J.C, & Lanier L.L. (2014). Proapoptotic Bim regulates antigen-specific NK cell contraction and the generation of the memory NK cell pool after cytomegalovirus infection. The Journal of Experimental Medicine, 211(7), 1289-1296.
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10

Cytotoxicity of Anti-FCRL5 Immunotoxins on B and T Cells

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The cytotoxic effects of the anti–FCRL-5 immunotoxins on purified B cells and T cells from HCV-infected patients with MC and healthy donors were measured by flow cytometry using 7-aminoactinomycin D (7-AAD) and annexin V staining. Prepared cell samples were seeded onto 96-well plates at 2 × 105 cells/well. Serial dilutions of F56-IT and F25-IT in the culture medium were added to the cells, at a final concentration of 0–10,000 ng/ml. After 72 hours of culture, the cells were stained with PE–Cy7–conjugated anti-CD19 or Alexa Fluor 700–conjugated anti-CD3 and FITC–conjugated anti-CD45 (Beckman Coulter), and then resuspended in annexin binding buffer and labeled with 7-AAD and PE-conjugated annexin V (BD PharMingen). Flow cytometric analyses were performed on a Navios flow cytometer, with results analyzed using Navios software (Beckman Coulter). Viability was assessed by setting the gates based on the light-scatter properties of the lymphocytes. Viable cells were those negative for 7-AAD and PE-conjugated annexin V. To compensate for spontaneous cell death, the relative percentage of viable cells at the end of the assay was calculated using the following formula: (no. of CD19+ or CD3+ viable cells recovered in the treated well/no. of CD19+ or CD3+ viable cells in the untreated well) × 100 (41 (link)). All experiments were performed in duplicate.
Terrier B., Nagata S., Ise T., Rosenzwajg M., Pastan I., Klatzmann D., Saadoun D, & Cacoub P. (2014). CD21−/low Marginal Zone B Cells Highly Express Fc Receptor–like 5 Protein and Are Killed by Anti–Fc Receptor–like 5 Immunotoxins in Hepatitis C Virus–Associated Mixed Cryoglobulinemia Vasculitis. Arthritis & rheumatology (Hoboken, N.J.), 66(2), 433-443.
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