Cells were washed with PBS, incubated with Human TruStain Fc Receptor Blocking Solution (BioLegend, 422302), and stained with the following fluorochrome-conjugated monoclonal antibodies: CD3-FITC (clone HIT3a), CD56-PE (clone HCD56), PD-1-APC (clone EH12.2H7), CD69-BV711 (clone FN50), Ki-67-PE-Cy7, and CD45-BV510 (clone HI30; all from BioLegend) and (clone 20Raj1, eBioscience). Live/dead staining was performed with Fixable Viability Dye 780 (eBioscience).
Cd45 bv510 clone hi30
Manufactured by BioLegend
CD45-BV510 (clone HI30) is a fluorochrome-conjugated monoclonal antibody that binds to the human CD45 antigen. CD45 is a leukocyte-specific receptor tyrosine phosphatase expressed on the surface of all hematopoietic cells.
Automatically generated - may contain errors
Lab products found in correlation
Fixable viability dye 780, by Thermo Fisher Scientific (3 mentions)
Human trustain fc receptor blocking solution, by BioLegend (2 mentions)
Lsrfortessa flow cytometer, by BD (2 mentions)
Cd56 pe clone hcd56, by BioLegend (2 mentions)
Cd3 fitc clone hit3a, by BioLegend (2 mentions)
Cd69 bv711 clone fn50, by BioLegend (1 mentions)
Ki 67 pe cy7, by BioLegend (1 mentions)
Influx cell sorter, by BD (1 mentions)
3 protocols using cd45 bv510 clone hi30
1
Multiparametric Immune Profiling
2
Flow Cytometric Analysis and Sorting of Immune Cells
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Human cells were washed with PBS and staining buffer, and then incubated with Human TruStain Fc receptor blocking solution (BioLegend, #422302) for 15 minutes. Cells were stained with the following fluorochrome-conjugated monoclonal antibodies: CD3-FITC (clone HIT3a, BioLegend), CD56-PE (clone HCD56, BioLegend), CD45-BV510 (clone HI30, BioLegend) and CD8-BV785 (clone RPA-T8, BioLegend). Live/dead staining was performed using Fixable Viability Dye 780 (eBioscience #65-0865-14). Data were acquired using a BD LSRFortessa flow cytometer (Becton Dickinson, San Jose, CA) and analyzed using FlowJo Software (Beckton Dickinson, San Jose, CA). Fluorescence-associated cell sorting (FACS) was performed using a BD inFlux cell sorter (Beckton Dickinson, San Jose, CA). Cells were sorted for blood and intra-tumoral NK cells (live CD45+CD3-CD56+), blood and intra-tumoral T cells (live CD45+CD3+CD56-), and tumor cells (live CD45-). Following FACS isolation, cells were cryopreserved and stored in liquid nitrogen for subsequent RNA extraction and sequencing.
3
Flow Cytometry of Blood and Tumor Samples
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Blood and tumor samples were processed for flow cytometry as described previously.25 31 32 (link) Cells were stained with the following fluorochrome-conjugated monoclonal antibodies: CD3-FITC (clone HIT3a, BioLegend), CD56-PE (clone HCD56, BioLegend), and CD45-BV510 (clone HI30, BioLegend). Live/dead staining was performed using Fixable Viability Dye 780 (eBioscience #65-0865-14). Flow cytometry results were acquired using a BD LSRFortessa flow cytometer (Becton Dickinson, San Jose, California, USA) and analyzed using FlowJo Software (Becton Dickinson, San Jose, California, USA).
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