MojoSort™ Mouse F4/80 Selection Kit

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Regulatory Status
RUO
Other Names
Hepatic Marcrophages Enrichment, Kupffer Cells Isolation, Peritoneal Macrophages Enrichment, Mouse Macrophages Isolation, F4/80 Macrophage Isolation, F4/80 Cell Isolation, Cell Separation, Macrophage Isolation Kit, F4/80 Positive Cells
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A_MojoSort_Mouse_F4slash80_Selection_Kit_A_122623
Figure 1. Cell suspensions from mouse liver non-parenchymal cells (LNPCs) were prepared for the positive isolation of F4/80+ cells using the MojoSort™ Mouse F4/80 Selection Kit. The cells were incubated with the biotin-antibody cocktail prior to the addition of the Streptavidin Nanobeads. Samples were collected before (panel 1a) and after magnetic separation (panel 1b) and stained with anti-CD11b (clone M1/70) APC (Cat. No. 101211) and anti-mouse F4/80 (clone W20065D) PE (Cat. No 111704). Cells were then further stained with anti-mouse CD45 (clone 30-F11) FITC (Cat. No. 103107) (panel 1c). Dead cells were excluded using Helix NP™ Blue (Cat. No. 425305) staining.
  • A_MojoSort_Mouse_F4slash80_Selection_Kit_A_122623
    Figure 1. Cell suspensions from mouse liver non-parenchymal cells (LNPCs) were prepared for the positive isolation of F4/80+ cells using the MojoSort™ Mouse F4/80 Selection Kit. The cells were incubated with the biotin-antibody cocktail prior to the addition of the Streptavidin Nanobeads. Samples were collected before (panel 1a) and after magnetic separation (panel 1b) and stained with anti-CD11b (clone M1/70) APC (Cat. No. 101211) and anti-mouse F4/80 (clone W20065D) PE (Cat. No 111704). Cells were then further stained with anti-mouse CD45 (clone 30-F11) FITC (Cat. No. 103107) (panel 1c). Dead cells were excluded using Helix NP™ Blue (Cat. No. 425305) staining.
  • B_MojoSort_Mouse_F4slash80_Selection_Kit_B_122623
    Figure 2. Cell suspensions from mouse peritoneal lavage were prepared for the positive isolation of F4/80+ cells using the MojoSort™ Mouse F4/80 Selection Kit. The cells were incubated with the biotin-antibody cocktail prior to the addition of the Streptavidin Nanobeads. Samples were collected before (panel 2a) and after magnetic separation (panel B) and stained with anti-CD11b (clone M1/70) APC (Cat. No. 101211) and anti-mouse F4/80 (clone W20065D) PE (Cat. No 111704). Dead cells were excluded using Helix NP™ Blue (Cat. No. 425305) staining.
  • C_MojoSort_Mouse_F4slash80_Selection_Kit_C_122723
    Figure 3. Cell suspensions from mouse liver non-parenchymal cells (LNPCs) were prepared for the positive isolation of F4/80+ cells using the MojoSort™ Mouse F4/80 Selection Kit and magnetic columns of cell separation. The cells were incubated with the biotin-antibody cocktail prior to the addition of the Streptavidin Nanobeads. Samples were collected before (panel 3a) and after column separation (panel 3b) and stained with anti-CD11b (clone M1/70) APC (Cat. No. 101211) and anti-mouse F4/80 (clone W20065D) PE (Cat. No 111704). Cells were then further stained with anti-mouse CD45 (clone 30-F11) FITC (Cat. No. 103107) (panel 3c). Dead cells were excluded using Helix NP™ Blue (Cat. No. 425305) staining.
  • D_MojoSort_Mouse_F4slash80_Selection_Kit_D_122623
    Figure 4. Cell suspensions from mouse peritoneal lavage were prepared for the positive isolation of F4/80+ cells using the MojoSort™ Mouse F4/80 Selection Kit and magnetic columns of cell separation. The cells were incubated with the biotin-antibody cocktail prior to the addition of the Streptavidin Nanobeads. Samples were collected before (panel 4a) and after column separation (panel 4b) and stained with anti-CD11b (clone M1/70) APC (Cat. No. 101211) and anti-mouse F4/80 (clone W20065D) PE (Cat. No 111704). Dead cells were excluded using Helix NP™ Blue (Cat. No. 425305) staining.
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480169 10 tests 160€
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480170 100 tests 400€
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Description

Mouse F4/80+ cells are either selected or depleted by incubating the sample with the Biotin anti-mouse F4/80 Antibody Cocktail followed by Streptavidin Nanobeads. The magnetically labeled fraction is retained by the use of a magnetic separator. The magnetically labeled fraction contains the cells of interest; do not discard this fraction. After collection of the F4/80+ expressing cells, 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. Please contact our technical service team for further assistance.

Product Details
Technical Data Sheet (pdf)

Kit Contents

Kit Contents

For Cat# 480169:

  • 1 vial containing 100 µL of anti-mouse F4/80 Biotin-Antibody Cocktail
  • 1 vial containing 100 µL of Streptavidin Nanobeads

For Cat# 480170:

  • 1 vial containing 1 mL of anti-mouse F4/80 Biotin-Antibody Cocktail
  • 1 vial containing 1 mL of Streptavidin Nanobeads

Product Details

Verified Reactivity
Mouse
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

From mouse peritoneal lavage:

  • 10 µL of Biotin-Antibody Cocktail for 2 x 106 peritoneal lavage cells in 100 µL of buffer.
  • 10 µL of Streptavidin Nanobeads for 2 x 106 peritoneal lavage cells in 100 µL of buffer.

From mouse liver:

  • 4 µL of Biotin-Antibody Cocktail for 1 x 107 liver nonparenchymal cells in 100 µL of buffer.
  • 3.2 µL of Streptavidin Nanobeads for 1 x 107 liver nonparenchymal cells in 100 µL of buffer.
Application Notes

This kit is designed for the positive selection of F4/80+ cells from mouse peritoneal lavage and liver.

Test sizes/starting cell capacity:

  • The 10 and 100 test sizes provide enough reagents for approximately 2 x 107 and 2 x 108 peritoneal lavage cells, respectively.
  • For liver cells, The 10 and 100 test sizes provide enough reagents for approximately 25 and 250 tests, respectively, due to lower volumes of reagents needed per test in this sample type. The 10 and 100 test sizes provide enough reagents for approximately 2.5 x 108 and 2.5 x 109 liver cells, respectively.

Optional:

  • To futher increase purity of F4/80+ cells from mouse liver, it is recommended to harvest liver nonparenchymal cells (LNPCs) from PBS-perfused tissue.
  • High yield of hepatic F4/80+ macrophages can be achieved by using magnetic columns.
  • The appropiate volume of antibody cocktail/nanobeads are critical and specific for distinct tissues.

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: 4X

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: 12.05.2023

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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