c-KIT-positive cells were enriched using a magnetic separation system (MACS) with anti-c-KIT magnetic beads (Miltenyi Biotec). The enriched cells were stained with c-KIT-APC (2B8, BioLegend), SCA-1-PE (E13–161.7, BioLegend), and lineage antibody cocktail (CD3, B220, CD11b, Gr-1, and Ter119; all from BD) conjugated with PE-Cy5. Dead cells were excluded using 7-amino-actinomycin-D staining. For the analysis of CXCR4 protein expression, c-KIT-enriched BM cells were stained with CXCR4-PE (2B11, eBioscience) or Isotype Control (eB149/10H5, eBioscience) together with CD48-FITC (HM48-1, BioLegend), CD150-PE-Cy7 (TC15-12F12.2, BioLegend), c-KIT-APC-eFluor780 (2B8, Fisher Scientific), SCA-1-Brilliant Violet 421 (D7, BioLegend), and lineage antibody cocktail conjugated with PE-Cy5. CD150+CD48− within LSK cells were defined as HSCs (Kiel et al., 2005 (link)). Cells were sorted on FACSAria III or FACSAria IIu and analyzed on FACS Fortessa, LSRII, or FACSCanto II (BD). Collected data were analyzed with FlowJo software (Tree Star).
Cxcr4 pe
Manufactured by Thermo Fisher Scientific
Sourced in United States
CXCR4-PE is a fluorescent-labeled antibody that binds to the CXCR4 chemokine receptor on the surface of cells. It is used in flow cytometry applications to detect and quantify CXCR4-expressing cells.
Automatically generated - may contain errors
Lab products found in correlation
Facscanto 2, by BD (1 mentions)
Facs fortessa, by BD (1 mentions)
C kit apc, by BioLegend (1 mentions)
Cd150 pe cy7, by BioLegend (1 mentions)
Anti c kit magnetic beads, by Miltenyi Biotec (1 mentions)
Lineage antibody cocktail, by BD (1 mentions)
Sca1 brilliant violet 421, by BioLegend (1 mentions)
Sca 1 pe, by BioLegend (1 mentions)
Fixation permeabilization solution kit, by BD (1 mentions)
Cd166 pe, by Thermo Fisher Scientific (1 mentions)
Cd133 2 pe, by Miltenyi Biotec (1 mentions)
Cd9 fitc, by Thermo Fisher Scientific (1 mentions)
Gr 1 fitc, by Thermo Fisher Scientific (1 mentions)
Ack lysing buffer, by Thermo Fisher Scientific (1 mentions)
Cd115 pe cy7, by Thermo Fisher Scientific (1 mentions)
Cd62l apc, by Thermo Fisher Scientific (1 mentions)
Facsaria 3, by BD (1 mentions)
Facsdiva v6, by BD (1 mentions)
Facs flow buffer, by BD (1 mentions)
Cd86 pe, by Thermo Fisher Scientific (1 mentions)
5 protocols using cxcr4 pe
1
Isolation and Analysis of Hematopoietic Stem Cells
2
Flow Cytometry Analysis of Cell Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Flow cytometry analysis of surface markers was performed as previously described [43 (link)]. Intracellular staining was performed by using Fixation/Permeabilization Solution Kit (BD Pharmingen), where cells were incubated with primary antibody over night, followed by washing and then incubated with secondary antibody for 2 h. Cells stained with secondary antibodies alone were used as control for gating. The following antibodies were used: CD166-PE, CD9- FITC, CXCR4-PE, m/hCD44-APC (eBioscience, San Deigo, CA), CD133/2-PE and anti-m/hAPC-A2B5 (Miltenyi, Billerica, MA).
3
Characterization of Aged Neutrophils
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Peripheral blood was lysed with ACK lysing buffer (Gibco, Cat# A1049201) and the nucleated cells were stained with the following conjugated monoclonal antibodies: CD62L-APC, CD115-PE-Cy7, CXCR4-PE, and Gr-1-FITC (all from eBioscience, USA). Flow cytometry was carried out on the Moflo XDP Sorter (BD Biosciences). Dead cells were excluded by FSC, SSC and Propidium iodide. Neutrophils were gated by Gr-1hi CD115lo SSChi and aged neutrophils gated by CD62Llo CXCR4hi within the neutrophil population.
4
Multiparameter Flow Cytometry Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Antibodies used for analysis were anti-human CD11c APC, CD80 FITC, CD83 FITC, HLA-DR FITC, CD40 PE, CD86 PE, TLR-2 PE, CXCR4 PE, CCR7 PE, CD4 PECy7, and IFN-γ APC (all from eBioscience). Cells were resuspended in PBS supplemented with fetal bovine serum (FBS) (HyClone Thermo Scientific, Logan, UT, USA), stained with specific antibodies, fixed with IC fixation buffer (eBioscience) and resuspended in FACSFlow buffer (Becton Dickinson, San Diego, CA, USA) for subsequent analysis. Data were acquired on a FACSAria III with FACSDiva v6.1.3 software (both Becton Dickinson) and analyzed by FlowJo software (Treestar, USA).
5
Flow Cytometry Analysis of CAR Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The following antibodies were used for flow cytometric analysis: EpCAM-APC (324208, BioLegend), mCD45-PE-CY7 (103114, BioLegend), hCD45-PE (555483, BD), CD45-PerCP-CY5.5 (558714, BD), CXCR4-PE (12–9999–42, eBioscience), p-mTOR Ser2448-PE (583489, BD), p-S6 Ser235/236-APC (14733S, CST), CD3-BV421(317344, BioLegend), and CD33-PE (555450, BD). Cells isolated from animals or cell lines cultured in vitro were washed once in PBS supplemented with 2% FBS (Gibco), and after blocking by Fc receptors, stained at 4°C for 30 minutes in the dark, washed twice with PBS, and then tested. For intracellular staining, cells were first fixed and permeabilized with Foxp3/Transcription Factor Staining Buffer (eBioscience) and then antibody staining was performed. The surface expression of EpCAM CAR was detected by staining with goat anti-mouse F(ab')2 antibody conjugated with Alexa Fluor 647 from Jackson ImmunoResearch. BD LSRII flow cytometry was used for detection, and FlowJo was used for analysis.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!