4T1, H22, HepG2, NIH‐3T3, and Panc02 were purchased from BeNa Culture Collection, China. Anti‐mouse PD‐1 antibody (PD‐1 Ab, BE0146‐100 mg) was purchased from Bio X Cell. Anti‐human PD‐1 Ab was obtained as a gift from Livzon Pharmaceutical Group Inc., China. Flow cytometry antibodies included PE anti‐mouse CD274 (B7‐H1, PD‐L1) (124308), APC/Cyanine7 anti‐mouse CD45 (103132), FITC anti‐mouse CD3 (100204), Brilliant Violet 421 anti‐mouse CD4 (100563), PE/Cy7 anti‐mouse CD8a (100722), PE anti‐mouse IFN‐γ (505808), Alexa Fluor 647 anti‐human/mouse Granzyme B (515406), APC anti‐mouse Ki‐67 (652406), PE anti‐mouse/human CD44 (103008), PE anti‐mouse Ly‐6G/Ly‐6C (Gr‐1) (108407), APC/Cyanine7 anti‐mouse/human CD11b (101226), Brilliant Violet 421 anti‐mouse F4/80 (123137), PE anti‐mouse CD86 (105008), PE/Cy7 anti‐mouse CD206 (MMR) (141720), Alexa Fluor 647 anti‐mouse FOXP3 (126408), APC anti‐mouse CD49b (pan‐NK cells) (108910), PE anti‐mouse CD25 (102008), PE anti‐mouse/human CD44 (103008), APC anti‐mouse CD62L (104412), Zombie Aqua Fixable Viability Kit (423102), Cell Activation Cocktail (with Brefeldin A) (423304), True‐Nuclear Transcription Factor Buffer Set (424401), and Intracellular Staining Permeabilization Wash Buffer (10X) (421002) were purchased from BioLegend.
Apc anti mouse ki 67
Manufactured by BioLegend
The APC anti-mouse Ki-67 is a flow cytometry reagent used for the detection and quantification of the Ki-67 protein, which is a cellular marker for proliferation. It is conjugated to the fluorescent dye allophycocyanin (APC) for detection.
Automatically generated - may contain errors
Lab products found in correlation
Hepg2, by BNCC (1 mentions)
Nih 3t3, by BNCC (1 mentions)
Panc02, by BNCC (1 mentions)
Apc cyanine7 anti mouse cd45, by BioLegend (1 mentions)
Fitc anti mouse cd3, by BioLegend (1 mentions)
Brilliant violet 421 anti mouse cd4, by BioLegend (1 mentions)
Pe cy7 anti mouse cd8a, by BioLegend (1 mentions)
Pe anti mouse human cd44, by BioLegend (1 mentions)
Brilliant violet 421 anti mouse f4 80, by BioLegend (1 mentions)
Pe anti mouse cd86, by BioLegend (1 mentions)
Alexa fluor 647 anti mouse foxp3, by BioLegend (1 mentions)
Apc anti mouse cd49b, by BioLegend (1 mentions)
Pe anti mouse cd25, by BioLegend (1 mentions)
Apc anti mouse cd62l, by BioLegend (1 mentions)
Intracellular staining permeabilization wash buffer 10x, by BioLegend (1 mentions)
True nuclear transcription factor buffer set, by BioLegend (1 mentions)
Pe anti mouse ifn γ, by BioLegend (1 mentions)
Zombie aqua fixable viability kit, by BioLegend (1 mentions)
Cell activation cocktail with brefeldin a, by BioLegend (1 mentions)
Fitc anti mouse cd11b, by BioLegend (1 mentions)
4 protocols using apc anti mouse ki 67
1
Comprehensive Immunological Profiling of Tumor Cell Lines
2
Analyzing Tumor-Associated Myeloid Cells
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The blood collected from the tumor-bearing mice was lysed with ammonium-chloride-potassium lysis buffer (cat. no. 00-4333-57; Thermo Fisher Scientific, Inc.) and washed with pre-cooled PBS to harvest cells by centrifugation (1,000 × g, 5 min at 4°C). The cells were incubated with PE anti-mouse Ly-6G/Ly-6C (Gr1; 0.25 μg/μl; cat. no. 108407; BioLegend, Inc.), FITC anti-mouse CD11b (0.25 μg/μl; cat. no. 101205; BioLegend, Inc.) and APC anti-mouse Ki-67 (0.25 μg/μl; cat. no. 652405; BioLegend, Inc.) antibodies for 40 min at 4°C. The positive cells were measured using a FACSCanto II Flow Cytometry System (BD Biosciences) and analyzed by FlowJo version 10 (FlowJo LLC).
3
Phenotypic and Functional Characterization of Stem Cells
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Cells were incubated with APC/Cy7 anti-mouse Ly-6A/E (Sca-1) (BioLegend, San Diego, CA, USA, 108125, dilution 1:200), FITC anti-mouse CD117 (c-Kit) (BioLegend, 105805; dilution 1:200), AlexaFluor488 anti-mouse PDGFRβ (Becton Dickinson, Heidelberg, Germany 558427, dilution 1:200), and Hoechst 33342 (Sigma Aldrich, Hamburg, Germany; dilution 1:10,000) for 1 h at 4 °C in the dark. For the proliferation analysis, MACS-sorted cells were fixed with chilled ethanol and incubated at 20 °C for 2 h followed by APC anti-mouse Ki67 (BioLegend, 652405, dilution 1:200) and Hoechst for 30 min at room temperature (RT) in the dark. For the apoptosis analysis, MACS-sorted cells were resuspended in annexin V binding buffer (BioLegend, 422201) and stained with Annexin V 647 (BioLegend, 640911, dilution 1:20) and Zombie Green (BioLegend, 423111, dilution 1:1000) for 10 min in the dark. Samples were washed twice with PBS and analyzed by flow cytometry (BD FACS Canto II and BD FACS Aria III).
4
Flow Cytometry Analysis of MSC Surface Markers
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Characterization of the surface markers of cells was performed by using flow cytometry. MSCs digested in a dish with 0.25% Trypsin (Biosharp) were suspended in staining buffer (PBS, 1% FBS) and 1 × 105 cells/sample were incubated with fluorescently labeled antibodies (all 1:200) for 30 min at 4 °C. The antibodies used for flow cytometry were as follows: PE anti-mouse CD29 (eBioscience), PE anti-mouse Sca-1 (Biolegend), PE anti-mouse CD140a (eBioscience), PE anti-mouse CD44 (eBioscience), PE anti-mouse CD73 (eBioscience), PE anti-mouse MHC II (eBioscience), PE anti-mouse CD45 (Biolegend), PE anti-mouse CD31 (eBioscience), PE anti-mouse CD34 (Biolegend), PE anti-mouse CD11B (eBioscience), PE anti-mouse ICAM-1 (Biolegend), and BV421 anti-mouse VCAM-1 (BD Pharmingen). For intracellular staining, cells stained with cell surface antibodies (PE anti-mouse CD3 (Biolegend)) were fixed, permeabilized using foxp3/transcription factor concentrate and diluent kit set prior to incubation with antibodies directed at intracellular antigens (APC anti-mouse KI-67 (Biolegend)). Fluorescence intensity was measured by flow cytometry (Cytoflex, Beckman Coulter). Data were analyzed with FlowJo v10 software and Cytexpert software.
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