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Protocols and Applications

Application

Access the Breadth of Sample Preparation Protocols to Awaken True Functional Immune Biology

Precedent Protocols and Processes ensure that enrichment, stimulation and labeling will result in maximizing the correlative data achieved, while minimizing non-specific impact on the cells themselves. Highlights: • Published enrichment, stimulation, and staining protocols throughout immunotherapy ... More


Protocol

Enrichment of transfected cells with Dharmacon Edit-R Fluorescent dCas9-VPR mRNA

Edit-R Fluorescent dCas9-VPR mRNA enables both transfection optimization and enrichment for gene modulation experiments. For optimal enrichment of edited cells, sorting the cells for high EGFP expression levels (selecting the top 10% fluorescence) in addition to negative and dim fluorescence for com... More


Protocol

Dharmacon Edit-R Lentiviral sgRNA glycerol stocks

The Dharmacon™ Edit-R™ Lentiviral sgRNA vector is part of the Edit-R CRISPR-Cas9 system for genome engineering. The purpose is to provide the researcher with the most effective tools to deliver a gene-specific sgRNA and, together with Cas9 expression, allow gene editing in cells. In addition to the ... More


Protocol

CRISPRmod dCas9-VPR or dCas9- SALL1-SDS3 mRNA and synthetic guide RNA transfection protocol

The following is a protocol for transfecting CRISPRmod dCas9-VPR or dCas9-SALL1-SDS3 mRNA with synthetic guide RNA into cultured mammalian cells using DharmaFECT™ Duo transfection reagent (Cat #T-2010-xx). The protocol is written for transfection into 96-well tissue culture plates.... More


Protocol

IsoCode Single-Cell Innate Immune: Human CD34+ Myeloid Stem Cell Protocol

A. Overview1. Overview of ProtocolDay 1: Cryopreserved cells are thawed and cultured overnight in the presence of IL-2.Day 2: Enrichment and Stimulation of CD34+ cells for 24 hours.Day 3: Staining and Loading of CD34+ cells onto IsoCode chip.NOTE: This protocol outlines the standard method for thawi... More


Protocol

IsoCode Single-Cell Innate Immune: Human Monocyte Protocol Using Membrane Stain

A. OverviewOverview of ProtocolDay 1: Cryopreserved cells are thawed and cultured overnight in complete RPMI media.Day 2: Enrichment and Stimulation of Monocytes for 24 hours.Day 3: Staining and Loading of Monocytes onto IsoCode Chip.... More


Protocol

IsoCode Single-Cell Adaptive Immune: TCR-T Protocol

A. Overview of ProtocolDay 1: Cryopreserved TCR-T cells are thawed and cultured overnight in the presence of IL-2. Cryopreserved target cell lines are thawed, cultured and passaged several days prior to beginning the experiment.Day 2: Enrichment, Staining, and Antigen Stimulation of CD4+ and/or CD8+... More


Protocol

USP Compendial Particle Measurements with the Aura System

The Aura platform provides a rapid and robust USP 788 compendial method for subvisible particle analysis, a critical quality attribute. It can handle a wide range of sample types and can analyze anywhere from 5 µL to 10 mL of sample, enabling continuity of method from developability assessment to lo... More


Protocol

USP Particle Count Standards with Aura and Particle Vue 3.1

All membrane microscopy methods must generate accurate and reproducible counts for bead counting standards. We present a validated methodology for accurately counting United States Pharmacopeia (USP) microsphere count standards on Aura systems by seamlessly calculating the cumulative area of all the... More


Protocol

Evaluate Cell Therapy Product Purity with Aura CL

The CAR-T development workflow is a multi-step process that includes transfection of T cells with a viral vector to express a chimeric antigen and activation/expansion of the CAR-T using a molecule like a Dynabead conjugated to CD3/CD28. Each of these steps is a potential source of contamination tha... More