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Pe cy7 anti human cxcr4

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PE-Cy7 anti-human CXCR4 is a fluorescent-labeled antibody that binds to the CXCR4 chemokine receptor on human cells. It can be used for the detection and analysis of CXCR4-expressing cells by flow cytometry.

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Lab products found in correlation

Fitc anti human cd15, by BioLegend (3 mentions) Annexin 5 fitc, by BioLegend (2 mentions) Celltiter glo luminescent viability assay, by Promega (2 mentions) Dexamethasone (dex), by Merck Group (2 mentions) Bortezomib, by Merck Group (2 mentions) Pomalidomide, by Merck Group (2 mentions) Bkt140, by Bachem (2 mentions) Pomalidomide, by Selleck Chemicals (2 mentions) Accuri c6 flow cytometer, by BD (2 mentions) Plerixafor, by Selleck Chemicals (2 mentions) Bortezomib, by Selleck Chemicals (2 mentions) P1585, by Merck Group (1 mentions) Facsverse, by BD (1 mentions) Annexin 5 pe 7 aad apoptosis kit, by MultiSciences Biotech (1 mentions) Lsrfortessa cell analyzer, by BD (1 mentions) Pe anti human cd63, by BioLegend (1 mentions)

Close spelling variants of this product

Anti-CXCR4 (13 citations) CXCR4 (9 citations) CXCR4-PE (8 citations) PE-anti-human CXCR4 (2 citations) Anti-CXCR4-PE (2 citations)

6 protocols using pe cy7 anti human cxcr4

1

Neutrophil ROS Production Assay

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For ROS analysis, 5 × 105 neutrophils were isolated from healthy controls and psoriasis patients and treated with phorbol 12-myristate 13-acetate (PMA, 50 nM, P1585, Sigma-Aldrich) at 37 °C for 30 min, then cells stained with FITC anti-human CD15 (301904, 1:100, BioLegend) and PE-Cy7 anti-human CXCR4 (306514, 1:100, BioLegend), and intracellular ROS production was measured by a dihydroethidium probe kit (BB-47051, DHE, BestBio, Beijing, China) at 37 °C for 30 min. After washing, the cells were immediately analyzed by flow cytometry.
Chen J., Bai Y., Xue K., Li Z., Zhu Z., Li Q., Yu C., Li B., Shen S., Qiao P., Li C., Luo Y., Qiao H., Dang E., Yin W., Gudjonsson J.E., Wang G, & Shao S. (2023). CREB1-driven CXCR4hi neutrophils promote skin inflammation in mouse models and human patients. Nature Communications, 14, 5894.
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2

Quantifying CXCR4 Expression in MSCs

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For the analysis of CXCR4 cell surface expression, MSCs were seeded at 5×104 cells per well of 6 well plates. The cells were treated or not for 24h with MDAMB231 CM. The cells were scrapped from the wells using rubber cell scrappers. For blocking, the cells were incubated in 1% FBS in PBS. For CXCR4 staining, PE/Cy7 anti-human CXCR4 (BioLegend) was used at 1:20 dilution in PBS 1% BSA. Cells were analyzed by FACSVerse (BD Bioscience). The controls, analysis gates were optimized for each cell line used according to standard practice. A minimum of 10,000 events were recorded for each sample and analysis was performed using FlowJo software.
Lourenco S., Teixeira V.H., Kalber T., Jose R.J., Floto R.A, & Janes S.M. (2015). Macrophage Migration Inhibitory Factor - CXCR4 is the dominant chemotactic axis in human mesenchymal stem cell recruitment to tumors. Journal of immunology (Baltimore, Md. : 1950), 194(7), 3463-3474.
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3

Modulating CXCR4 Expression in Multiple Myeloma

Cited 1 time
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MM cell lines were treated for 72h with pomalidomide (Selleck), bortezomib (Selleck), and dexamethasone (MilliporeSigma) individually or in combination (half IC10 concentration for bortezomib and half IC10 for MM1S for dexamethasone and pomalidomide, half IC50 for OPM2 and MOLP2) at the indicated concentrations. To investigate the effects of drug removal on CXCR4 expression, cells were treated for 72h, washed, and incubated in RPMI-1640 media without drug for a further 72h. Cells were stained with PE/Cy7 anti-human CXCR4 (12G5, BioLegend, 1:100) and FITC Annexin V (BioLegend) and were analyzed using a BD Accuri C6 Flow Cytometer. The mean fluorescence index of each sample was calculated using FlowJo software v10.
For bulk ATAC sequencing, MMCLs were treated with inhibitors for 72h and cells were ficolled. 50,000 cells were pelleted and ATAC sequencing was performed using the Omni-ATAC protocol as described by Corces et al.42 (link).
To assess sensitivity to CXCR4 inhibitors following pre-treatment with PVD, CXCR4 inhibitors plerixafor (Selleck) and BKT140 (Bachem) were added at IC50 concentrations after 72h. Cell viability was measured after a further 72h using the CellTiterGlo Luminescent Viability Assay (Promega).
Frede J., Anand P., Sotudeh N., Pinto R.A., Nair M.S., Stuart H., Yee A.J., Vijaykumar T., Waldschmidt J.M., Potdar S., Kloeber J.A., Kokkalis A., Dimitrova V., Mann M., Laubach J.P., Richardson P.G., Anderson K.C., Raje N.S., Knoechel B, & Lohr J.G. (2021). Dynamic transcriptional reprogramming leads to immunotherapeutic vulnerabilities in myeloma. Nature cell biology, 23(11), 1199-1211.
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4

Neutrophil Apoptosis Profiling by Flow Cytometry

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Viability assays were performed using the Annexin V-PE/7-AAD apoptosis kit (AP104, MultiSciences Biotech Co., Ltd.) following the manufacturer’s instructions. Neutrophils were resuspended in RPMI Medium 1640 (C11875500BT, Gibco, USA) containing 10% FBS at a concentration of 5 × 105 cells/mL and incubated for 24 h at 37 °C. Before and after incubation, cells were co-stained with FITC anti-human CD15 (301904, 1:100, BioLegend) and PE-Cy7 anti-human CXCR4 (306514, 1:100, BioLegend) for 30 min. After washing, cells were incubated with Annexin V (5 µL) and 7-AAD antibodies (10 µL) in 1 × Annexin V binding buffer for 5 min at room temperature and immediately analyzed by flow cytometry. Four cellular populations were distinguished: viable cells (Annexin V-PE and 7-AAD double-negative), early apoptotic cells (Annexin V-PE positive and 7-AAD negative), late apoptotic cells (Annexin V-PE and 7-AAD double-positive) and necrotic cells/cellular debris (Annexin V-PE negative and 7-AAD positive).
Chen J., Bai Y., Xue K., Li Z., Zhu Z., Li Q., Yu C., Li B., Shen S., Qiao P., Li C., Luo Y., Qiao H., Dang E., Yin W., Gudjonsson J.E., Wang G, & Shao S. (2023). CREB1-driven CXCR4hi neutrophils promote skin inflammation in mouse models and human patients. Nature Communications, 14, 5894.
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5

Neutrophil Degranulation Analysis in Psoriasis

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Degranulation of neutrophils was assessed by monitoring the cell surface expression of CD63. Blood samples obtained from healthy controls and psoriasis patients were collected and red blood cells were removed using Red Blood Lysing Buffer (FXP001, 4 A Biotech Co., Ltd). Then cells were labeled with FITC anti-human CD15 (301904, 1:100, BioLegend), PE-Cy7 anti-human CXCR4 (306514, 1:100, BioLegend) and PE anti-human CD63 (353004, 1:100, BioLegend) at 4 °C for 30 min. After three washes, cells were resuspended in PBS and analyzed using flow cytometry (649225, BD LSRFortessa Cell Analyzer), Background fluorescence levels were determined by Fluorescence Minus One (FMO).
Chen J., Bai Y., Xue K., Li Z., Zhu Z., Li Q., Yu C., Li B., Shen S., Qiao P., Li C., Luo Y., Qiao H., Dang E., Yin W., Gudjonsson J.E., Wang G, & Shao S. (2023). CREB1-driven CXCR4hi neutrophils promote skin inflammation in mouse models and human patients. Nature Communications, 14, 5894.
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6

Modulating CXCR4 Expression in Multiple Myeloma

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
MM cell lines were treated for 72h with pomalidomide (Selleck), bortezomib (Selleck), and dexamethasone (MilliporeSigma) individually or in combination (half IC10 concentration for bortezomib and half IC10 for MM1S for dexamethasone and pomalidomide, half IC50 for OPM2 and MOLP2) at the indicated concentrations. To investigate the effects of drug removal on CXCR4 expression, cells were treated for 72h, washed, and incubated in RPMI-1640 media without drug for a further 72h. Cells were stained with PE/Cy7 anti-human CXCR4 (12G5, BioLegend, 1:100) and FITC Annexin V (BioLegend) and were analyzed using a BD Accuri C6 Flow Cytometer. The mean fluorescence index of each sample was calculated using FlowJo software v10.
For bulk ATAC sequencing, MMCLs were treated with inhibitors for 72h and cells were ficolled. 50,000 cells were pelleted and ATAC sequencing was performed using the Omni-ATAC protocol as described by Corces et al.42 (link).
To assess sensitivity to CXCR4 inhibitors following pre-treatment with PVD, CXCR4 inhibitors plerixafor (Selleck) and BKT140 (Bachem) were added at IC50 concentrations after 72h. Cell viability was measured after a further 72h using the CellTiterGlo Luminescent Viability Assay (Promega).
Frede J., Anand P., Sotudeh N., Pinto R.A., Nair M.S., Stuart H., Yee A.J., Vijaykumar T., Waldschmidt J.M., Potdar S., Kloeber J.A., Kokkalis A., Dimitrova V., Mann M., Laubach J.P., Richardson P.G., Anderson K.C., Raje N.S., Knoechel B, & Lohr J.G. (2021). Dynamic transcriptional reprogramming leads to immunotherapeutic vulnerabilities in myeloma. Nature cell biology, 23(11), 1199-1211.
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