Cells were dissociated into single cells by Accutase (Thermo Fisher Scientific) and then fixed using 4% paraformaldehyde for 15 min at room temperature. Blocking and permeabilizing procedures referred to Zhao et al. protocol [16 (link)]. After the above processes, cells were suspended with 100 μL PBS containing 0.1% BSA and stained with PLZF monoclonal antibody-PE (1:200, Invitrogen, Mags.21F7) for 40 min under room temperature. Cells were then washed and resuspended with PBS containing 0.1% BSA, and the percentage of PLZF+ cells was detected by a DxFLEX flow cytometer (Beckman Coulter). For the detection of haploid cells, single cells were suspended with 300 μL cold PBS containing 10% FBS and then added with 700 μL cold ethanol. After stored at -20℃ overnight, cells were resuspended with PBS containing 100 μg/mL PI (Sigma Aldrich), 100 ng/mL RNase (CWBio, China) and 0.1% Triton X-100 for 20 min at room temperature. Cells were then washed and resuspended with PBS, and the percentage of haploid cells was detected by a DxFLEX flow cytometer. For the detection of apoptotic cells, single cells were stained with PI and Annexin V using a Annexin V-EGFP Apoptosis Detection Kit (KeyGEN BioTECH, China), and the percentage of apoptotic cells was detected by a DxFLEX flow cytometer.
Plzf monoclonal antibody pe
Manufactured by Thermo Fisher Scientific
The PLZF monoclonal antibody-PE is a laboratory reagent used for the detection and analysis of PLZF (Promyelocytic Leukemia Zinc Finger) protein in biological samples. It is a fluorescently-labeled antibody that binds specifically to the PLZF protein, allowing for its identification and quantification using flow cytometry or other immunodetection techniques.
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Lab products found in correlation
3 protocols using plzf monoclonal antibody pe
1
Flow Cytometric Analysis of Cellular States
2
Characterizing hPGCLCs and hESCs
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To analyze hPGCLCs or hESCs using cell surface markers, we stained the dissociated cells with CD24, CD90, EpCAM, and INTEGRINα6. Intracellular staining was performed using a BD kit (BD, 560589) according to the manufacturer’s instructions. For analysis, SSCLCs cells were dissociated into single cells using Accutase and fixed with 4% paraformaldehyde for 15 min at room temperature. After blocking and permeabilizing, the cells were suspended in 100μL of PBS containing 0.1% BSA and stained with a PLZF monoclonal antibody-PE (Invitrogen, 12-9320-82) for 40 min at room temperature. The cells were then washed, resuspended in PBS, and the percentage of PLZF-positive cells was detected by a DxFLEX flow cytometer (Beckman Coulter). The primary antibodies used in this study are listed in Supplementary Tables.
3
Single-Cell Flow Cytometry Analysis
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Cells were dissociated into single cells by Accutase (Thermo Fisher Scienti c) and then xed using 4% paraformaldehyde for 15 min at room temperature. Blocking and permeabilizing procedures referred to Zhao et al. protocol [16] . After the above processes, cells were suspended with 100 μL PBS containing 0.1% BSA and stained with PLZF monoclonal antibody-PE (1:200, Invitrogen, Mags.21F7) for 40 min under room temperature. Cells were then washed and resuspended with PBS containing 0.1% BSA, and the percentage of PLZF + cells was detected by a DxFLEX ow cytometer (Beckman Coulter). For the detection of haploid cells, single cells were suspended with 300 μL cold PBS containing 10% FBS and then added with 700 μL cold ethanol. After stored at -20℃ overnight, cells were resuspended with PBS containing 100 μg/mL PI (Sigma Aldrich), 100 ng/mL RNase (CWBio, China) and 0.1% Triton X-100 for 20 min at room temperature. Cells were then washed and resuspended with PBS, and the percentage of haploid cells were detected by a DxFLEX ow cytometer. For the detection of apoptotic cells, single cells were stained with PI and Annexin V using a Annexin V-EGFP Apoptosis Detection Kit (KeyGEN BioTECH, China), and the percentage of apoptotic cells were detected by a DxFLEX ow cytometer.
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