All flow cytometry was performed on a FACSCanto II platform or BD FACS Aria III. The data were analyzed using FlowJo software (BD) and the positive percentages were compared to fluorescence minus one (FMO) controls. To determine the expression of sLex on the surface of the cells, either rhFTVI-treated or buffer treated (negative control) HSC populations were placed in 96-well-plates, stained with 10 μg/mL anti-sLex antibody for 30 minutes at 4°C (HECA-452), resuspended in FACS buffer (10-mM EDTA, 5% FBS and HBSS) and washed twice with FACS buffer (200 μL/well). To detect E-selectin binding, 10 μg/mL of recombinant E-selectin human Ig (E-Ig) chimera was prepared in buffer (20 mM HEPES pH 7.5, 2 mM CaCl2 and 5% FBS) and used to stain cells. A PE-conjugated anti-human IgG Fc (1:200 dilution in chimera buffer; BioLegend) secondary antibody was used to detect E-Ig. As a control for E-selectin staining, 20-mM EDTA was added to the chimera buffer. To detect cell surface proteins, the HSCs populations were stained with 10 μg/mL of primary conjugated antibodies against surface markers CXCR4, CD49e, CD49d, and CD29 in 100 μL FACS buffer (2 mM EDTA, 5% FBS and HBSS) for 30 minutes at 4°C, washed three times with FACS buffer, and analyzing surface-marker expression.
Pe conjugated anti human igg fc
Manufactured by BioLegend
Sourced in United States
PE-conjugated anti-human IgG Fc is a laboratory reagent that binds to the Fc region of human immunoglobulin G (IgG). The PE (phycoerythrin) fluorescent dye is conjugated to the anti-human IgG Fc antibody, allowing for detection and quantification of IgG in various experimental applications.
Automatically generated - may contain errors
Lab products found in correlation
Facscanto 2 platform, by BD (1 mentions)
Facsaria 3, by BD (1 mentions)
Compensation beads, by Thermo Fisher Scientific (1 mentions)
Cd133 pe antibody, by Miltenyi Biotec (1 mentions)
Cytoflex flow cytometer, by Beckman Coulter (1 mentions)
Anti human egfr clone ay13, by BioLegend (1 mentions)
Anti human cd63 antibody clone h5c6, by BioLegend (1 mentions)
Human trustain fcx, by BioLegend (1 mentions)
Pe conjugated anti dykddddk, by BioLegend (1 mentions)
Lsrfortessa, by BD (1 mentions)
4 protocols using pe conjugated anti human igg fc
1
Flow Cytometric Analysis of HSC Markers
2
Identifying GBM Stem Cell Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Primary cultured GBM speroid cells were seeded at 5 × 105 cells per ultra-low attached TC culture dish and incubated at 37°C in humidified 5% CO2 for 24 hours. Next, trypsinized cells were stained with PE-CD133 antibody (130-113-108, Miltenyi Biotec, Bergisch Gladbach, Germany), or APC-SOX2 antibody (IC2018 A, R&D systems, Minneapolis, MN, USA) for 60 minutes at room temperature. Control groups were unstained samples, or those incubated with PE-conjugated anti-human IgG Fc (410707, BioLegend), with or without compensation beads (01-3333-42, Thermo Fisher Scientific). A gated positive signal corresponding to PE-CD133 or APC-SOX2 antibodies was used with compensation beads. The cell sample was analyzed using flow cytometry (CytoFlex flow cytometer, Beckman Coulter, Indianapolis, IN, USA).
3
Intact EV Labeling and Characterization
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Whole conditioned media, from Section 2.3 , was taken directly for intact EV labelling. Conditioned media was aliquoted into 100 μl aliquots in replicates of 3 for each labelling condition (n = 3). For antibody labelling, EVs were first blocked using Human TruStain FcX (BioLegend, San Diego, CA, USA) for 10 min at RT. PE‐conjugated anti‐human antibody was added according to the optimized volumes. The anti‐human CD63 antibody (Clone H5C6; BioLegend, San Diego, CA, USA), anti‐human CD9 antibody (Clone H19a; BioLegend, San Diego, CA, USA), and anti‐human EGFR (Clone AY13; BioLegend, San Diego, CA, USA) antibodies were used for this study. IgG controls were performed using the PE‐conjugated anti‐human IgG Fc (Clone M1310G05; BioLegend, San Diego, CA, USA). CFDA‐SE labelling was performed using a 1:100 dilution of 10 mM CFDA‐SE (CFDA; Invitrogen, Waltham, MA) in dimethyl sulfoxide (DMSO; Sigma Aldrich, St. Louis, MO). Following labelling, unbound antibody and dye was eliminated via 100 kDa centrifugation filtration. The final sample was restored to 100 μl using 0.22 μm filtered 1X PBS.
4
CD19 and PD-L1 Expression Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For tumor cells expressing CD19 and/or PD-L1, 5 × 105 tumor cells were harvested and washed twice with FACS buffer (1× PBS containing 2% FBS). Then, tumor cells were stained with 0.5 μL of APC/Cy7-conjugated mouse anti-human CD19 (BD Pharmingen #557791) or 0.5 μL of APC-conjugated mouse anti-human PD-L1 (eBioscience #17–5983-73) at 4 °C for 30 min, washed with FACS buffer twice, and resuspended in FACS buffer for assessment. Additionally, 5 × 105 washed tumor cells were incubated with 2 μg/mL of trastuzumab prepared in our laboratory at 4 °C for 30 min, washed twice with FACS buffer, and further stained with 0.5 μL of PE-conjugated anti-human IgG Fc (Biolegend #409304) at 4 °C for 30 min, washed with FACS buffer twice, and then resuspended in FACS buffer to detect HER2. To detect the HER2 CAR, PD-L1 CAR or PD-L1 CCR presented on the T cells surface, T cells were stained with Alexa Fluor 647-conjugated anti-Myc tag (CST #2233S) or PE-conjugated anti-DYKDDDDK (Biolegend #637310). T cells (5 × 105) were harvested and washed twice with FACS buffer. For Myc or Flag tag staining, T cells were stained with 0.5 μL of Alexa Fluor 647-conjugated anti-Myc tag or PE-anti-DYKDDDDK at 4 °C for 30 min, washed twice with FACS buffer, and then resuspended in FACS buffer for detection. Fluorescence was assayed using a BD LSRFortessa, and all FACS data were analyzed with FlowJo V10 software.
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!