MojoSort™ Human Pan DC Isolation Kit

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Regulatory Status
RUO
Other Names
Dendritic Cell, Dendritic Cell Separation, Dendritic Cell Enrichment, DC Enrichment
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MojoSort_Human_Pan_DC_Isolation_Kit_1_120423
FIGURE 1. FLOW CYTOMETRIC ANALYSIS AFTER MAGNETIC SEPARATION. A single cell suspension from human PBMCs was prepared for the isolation of pan-dendritic cells using the MojoSort™ Human Pan DC Isolation Kit. The cells were incubated with Human TruStain FcX™ and the biotin-antibody cocktail prior to the addition of the Streptavidin Nanobeads. The cells were then subject to two rounds of magnetic separation. Then, the isolated cells were stained for CD45 (clone HI30) PE/Dazzle™ 594, Lineage cocktail [CD3 (clone SK7), CD15 (clone HCD14), CD19 (clone SJ25C1), CD20 (clone 2H7), CD34 (clone 561), CD56 (clone HCD56)] Alexa Fluor® 700, CD303 (clone 201A) Brilliant Violet 421™, CD1c (clone L161) PE, CD141 (clone M80) APC, CD303 (clone 201A) Brilliant Violet 421™, CD16 (clone 3G8) FITC and CD11c (clone Bu15) APC/Cyanine7. T cells, B cells, monocytes, progenitor cells, NK cells, and dead cells were excluded by using CD3, CD20, CD19, CD14, CD34, CD56, and 7-AAD expression. Within the total leukocyte population (panel A), purity of the isolated pan DCs can be determined by combining the DC subpopulations (panels B-D) and typically ranges between 70-90% depending on donors.
  • MojoSort_Human_Pan_DC_Isolation_Kit_1_120423
    FIGURE 1. FLOW CYTOMETRIC ANALYSIS AFTER MAGNETIC SEPARATION. A single cell suspension from human PBMCs was prepared for the isolation of pan-dendritic cells using the MojoSort™ Human Pan DC Isolation Kit. The cells were incubated with Human TruStain FcX™ and the biotin-antibody cocktail prior to the addition of the Streptavidin Nanobeads. The cells were then subject to two rounds of magnetic separation. Then, the isolated cells were stained for CD45 (clone HI30) PE/Dazzle™ 594, Lineage cocktail [CD3 (clone SK7), CD15 (clone HCD14), CD19 (clone SJ25C1), CD20 (clone 2H7), CD34 (clone 561), CD56 (clone HCD56)] Alexa Fluor® 700, CD303 (clone 201A) Brilliant Violet 421™, CD1c (clone L161) PE, CD141 (clone M80) APC, CD303 (clone 201A) Brilliant Violet 421™, CD16 (clone 3G8) FITC and CD11c (clone Bu15) APC/Cyanine7. T cells, B cells, monocytes, progenitor cells, NK cells, and dead cells were excluded by using CD3, CD20, CD19, CD14, CD34, CD56, and 7-AAD expression. Within the total leukocyte population (panel A), purity of the isolated pan DCs can be determined by combining the DC subpopulations (panels B-D) and typically ranges between 70-90% depending on donors.
  • MojoSort_Human_Pan_DC_Isolation_Kit_2_120423
    Figure 2. FLOW CYTOMETRIC ANALYSIS BEFORE MAGNETIC SEPARATION (CONTROL FOR FIG 1). A single cell suspension from human PBMCs was prepared for the isolation of pan-dendritic cells using the MojoSort™ Human Pan DC Isolation Kit. The cells were incubated with Human TruStain FcX™ and the biotin-antibody cocktail prior to the addition of the Streptavidin Nanobeads. As a control, a sample was collected before magnetic separation. Then, the cells were stained for CD45 (clone HI30) PE/Dazzle™ 594, Lineage cocktail [CD3 (clone SK7), CD15 (clone HCD14), CD19 (clone SJ25C1), CD20 (clone 2H7), CD34 (clone 561), CD56 (clone HCD56)] Alexa Fluor® 700, CD303 (clone 201A) Brilliant Violet 421™, CD1c (clone L161) PE, CD141 (clone M80) APC, CD303 (clone 201A) Brilliant Violet 421™, CD16 (clone 3G8) FITC and CD11c (Bu15) APC/Cyanine7. T cells, B cells, monocytes, progenitor cells, NK cells, and dead cells were excluded by using CD3, CD20, CD19, CD14, CD34, CD56, and 7-AAD expression. Within the total leukocyte population (panel A), purity of the isolated pan DCs can be determined by combining the DC subpopulations (panels B-D) and typically ranges between 70-90% depending on donors.
  • MojoSort_Human_Pan_DC_Isolation_Kit_3_120423
    Figure 3. FLOW CYTOMETRIC ANALYSIS BEFORE COLUMN SEPARATION. A single cell suspension from human PBMCs was prepared for the isolation of pan-dendritic cells using the MojoSort™ Human Pan DC Isolation Kit and LS Columns (Miltenyi Biotec). The cells were incubated with Human TruStain FcX™ and the biotin-antibody cocktail prior to the addition of the Streptavidin Nanobeads. The cells were then applied to the column for separation. Then, the isolated cells were stained for CD45 (clone HI30) PE/Dazzle™ 594, Lineage cocktail [CD3 (clone SK7), CD15 (clone HCD14), CD19 (clone SJ25C1), CD20 (clone 2H7), CD34 (clone 561), CD56 (clone HCD56)] Alexa Fluor® 700, CD303 (clone 201A) Brilliant Violet 421™, CD1c (clone L161) PE, CD141 (clone M80) APC, CD303 (clone 201A) Brilliant Violet 421™, CD16 (clone 3G8) FITC and CD11c (clone Bu15) APC/Cyanine7. T cells, B cells, monocytes, progenitor cells, NK cells, and dead cells were excluded by using CD3, CD20, CD19, CD14, CD34, CD56, and 7-AAD expression. Within the total leukocyte population (panel A), purity of the isolated pan DCs can be determined by combining the DC subpopulations (panels B-D) and typically ranges between 70-90% depending on donors.
  • MojoSort_Human_Pan_DC_Isolation_Kit_4_120423
    Figure 4. FLOW CYTOMETRIC ANALYSIS BEFORE COLUMN SEPARATION (CONTROL FOR FIG 3). A single cell suspension from human PBMCs was prepared for the isolation of pan-dendritic cells using the MojoSort™ Human Pan DC Isolation Kit and LS Columns (Miltenyi Biotec). The cells were incubated with Human TruStain FcX™ and the biotin-antibody cocktail prior to the addition of the Streptavidin Nanobeads. As a control, a sample was collected before separation. Then, the cells were stained for CD45 (clone HI30) PE/Dazzle™ 594, Lineage cocktail [CD3 (clone SK7), CD15 (clone HCD14), CD19 (clone SJ25C1), CD20 (clone 2H7), CD34 (clone 561), CD56 (clone HCD56)] Alexa Fluor® 700, CD303 (clone 201A) Brilliant Violet 421™, CD1c (clone L161) PE, CD141 (clone M80) APC, CD303 (clone 201A) Brilliant Violet 421™, CD16 (clone 3G8) FITC and CD11c (clone Bu15) APC/Cyanine7. T cells, B cells, monocytes, progenitor cells, NK cells, and dead cells were excluded by using CD3, CD20, CD19, CD14, CD34, CD56, and 7-AAD expression. Within the total leukocyte population (panel A), purity of the isolated pan DCs can be determined by combining the DC subpopulations (panels B-D) and typically ranges between 70-90% depending on donors.
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480143 10 tests 224€
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480144 100 tests 600€
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Description

Human cells which are non pan dendritic cells (Pan DC) are depleted by incubating the sample with a biotin antibody cocktail followed by incubation with magnetic Streptavidin Nanobeads. The magnetically labeled fraction is retained by the use of a magnetic separator. The untouched plasmacytoid dendritic cells (PDCs) and myeloid dendritic cells (MDCs) are collected by decanting the liquid into a clean tube. These are the cells of interest; do not discard the liquid. Some of the downstream applications include functional assays, gene expression, phenotypic characterization, etc.

MojoSort™ reagents are also compatible with column-based cell separation systems available from other vendors. Optimized protocols for cell separation using columns from in-house testing are provided for each kit under the “Related Protocols” section, as well as representative data on the product webpage (where available). Data generated using column separators are indicated on the figure legend.

Due to the property of the beads, MojoSort™ reagents typically require dilution for optimal use on column separators. Where available, recommended dilution factors for each kit component based on in-house testing are provided under the “Application Notes” section of the webpage.

Product Details
Technical Data Sheet (pdf)

Kit Contents

Kit Contents

For Cat# 480143:

  • 1 vial containing 250 μL of Human TruStain FcX™
  • 1 vial containing 100 µL of Human Pan DC Biotin-Antibody Cocktail
  • 1 vial containing 100 µL of Streptavidin Nanobeads

For Cat# 480144:

  • 1 vial containing 1 mL of Human TruStain FcX™
  • 1 vial containing 1 mL of Human Pan DC Biotin-Antibody Cocktail
  • 1 vial containing 1 mL of Streptavidin Nanobeads

Product Details

Verified Reactivity
Human
Formulation
Cocktail: Phosphate buffer solution containing 0.09% sodium azide, 0.5% BSA, 5% HPCD, pH 7.2.

Particle: Aqueous solution containing BSA and 0.05% sodium azide.
Preparation
The antibodies were purified by affinity chromatography, and conjugated with biotin under optimal conditions. The solution is free of unconjugated biotin.

Streptavidin Nanobeads: Streptavidin-coated magnetic beads.
Storage & Handling
All components should be stored undiluted between 2°C and 8°C.
Application

Cell Separation (MojoSort™) - Quality tested

Recommended Usage

10 µL of Human TruStain FcX™ for 1 x 107 cells in 100 µL of buffer.
10 µL of Biotin-Antibody Cocktail for 1 x 107 cells in 100 µL of buffer.
10 µL of Streptavidin Nanobeads for 1 x 107 cells in 100 µL of buffer.

Application Notes

This kit is designed for the isolation of untouched pan dendritic cells (pan DCs) from PBMCs. Purity of the isolated Pan DCs ranges between 70-90% within the total leukocyte population and is donor-dependent.

​Optional: To further enrich the dendritic cells, an optional second round of separation can be done to increase DC purity (see protocol). This kit provides enough reagents for 1 round of separation per test for the specified kit size. If performing a second round of separation with this kit, the 10 and 100 tests sizes provide enough reagents for approximately 9 and 90 tests, respectively.

Each lot has been individually optimized. Do not mix and match components from different lots or different kits.

Antibody or cocktail dilution to use in column: 1X
Nanobead dilution to use in column: 2X

Antigen Details

Cell Type
Dendritic cells
Biology Area
Adaptive Immunity, Innate Immunity
Gene ID
NA
UniProt
View information about Pan DC on UniProt.org

Related FAQs

Is there a way to detach your magnetic particles from the cell surface?

MojoSort™ Nanobeads: These magnetic particles cannot be removed from cells. This includes standalone Nanobeads and MojoSort kits using Nanobeads. We have found that cells are functional without the need to detach the magnetic Nanobeads.

 

MojoSort Microbeads: These magnetic particles may be released following the detailed product-specific protocol.

What is the size of your magnetic particles?

MojoSort Nanobeads: the average diameter is approximately 130 nm.

 

MojoSort Microbeads: the average diameter is approximately 1 μm.

Are MojoSort™ Nanobeads compatible with other commercially available magnetic separation systems?

MojoSort™ magnetic particles can be used with other commercially available magnetic separators, both free standing magnets and column-based systems. Because MojoSort™ protocols are optimized for the MojoSort™ separator, the protocols may need to be adjusted for other systems.  Please contact BioLegend Technical Service for more product-specific information and guidance. We do not recommend using MojoSort™ particles for BD’s IMag™ or Life Technologies’ DynaMag™.

What antibodies are present in the depletion cocktails provided for isolation kits?

Please contact our technical service team for further assistance.

Go To Top Version: 1    Revision Date: 11-28-2023

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

BioLegend, the BioLegend logo, and all other trademarks are property of BioLegend, Inc. or their respective owners, and all rights are reserved.

 

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Toll-Free Phone: 1-877-Bio-Legend (246-5343) Phone: (858) 768-5800 Fax: (877) 455-9587

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