Mouse Erythrocyte Lysing Kit

Catalog # Availability Size / Price Qty
WL2000
Product Details
Procedure
Citations (21)
FAQs
Reviews (3)

Mouse Erythrocyte Lysing Kit Summary

Kit Summary

For the removal of erythrocytes (RBC) from splenocyte preparations.

Key Benefits

  • Gently lyses erythrocytes
  • Stabilizes leukocytes for analysis by flow cytometry
  • Maintains natural light scattering and fluorescent staining properties of leukocytes
  • Maintains viability of leukocytes for subsequent tissue culture manipulation

Why Remove Erythrocytes?

The lysis of erythrocytes from whole blood is an important initial step in the isolation and analysis of enriched leukocyte preparations. Recovered immune cells can be accurately characterized following red blood cell removal. Lysis of erythrocytes under conditions that do not disrupt lymphocytes or myeloid cells is critical for downstream applications utilizing leukocytes harvested from whole blood.

Reagents Provided

The Mouse Erythrocyte Lysing Kit (Catalog # WL2000) contains the following reagents for the lysis of erythrocytes in splenocyte preparations:

  • M-Lyse Buffer Concentrate (10X)
  • Wash Buffer Concentrate (10X)
  • Fixative Concentrate (10X)

*This kit contains sufficient reagents to process 12.5 x 109 splenocytes.

Stability and Storage

Store all reagents at 20 °C to 25 °C. 

Specifications

Shipping Conditions
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Storage
Store the unopened product at room temperature. Do not use past expiration date.
Species
Mouse

Product Datasheets

⚠ WARNING: This product can expose you to chemicals including formaldehyde and methanol, which are known to the State of California to cause cancer and reproductive toxicity with developmental effects. For more information, go to www.P65Warnings.ca.gov.

Assay Procedure

Refer to the product datasheet for complete product details.

Briefly, erythrocytes can be lysed in splenocyte preparations using the following procedure:

  • Stain splenocytes or an enriched mononuclear cell preparation with antibody
  • Add M-Lyse Buffer
  • Wash cells with Wash Buffer
  • Fix cells for flow cytometry with Fixative
 

Reagents Provided

The Mouse Erythrocyte Lysing Kit (Catalog # WL2000) contains the following reagents for the lysis of erythrocytes in splenocyte preparations:

  • M-Lyse Buffer Concentrate (10X)
  • Wash Buffer Concentrate (10X)
  • Fixative Concentrate (10X)

This kit contains sufficient reagents to process 12.5 x 109 splenocytes.

Other Supplies Required

  • Ficoll-Hypaque™
  • Sterile distilled or deionized water
  • Sterile centrifuge tubes
  • Benchtop centrifuge
  • Pipettes and sterile pipette tips

Procedure Overview

R&D Systems Protocol for the Lysis of Erythrocytes Using the Mouse Erythrocyte Lysing Kit

 
  1. Stain a single cell suspension of mononuclear cells from a mouse spleen with antibody or antibodies (if performing flow cytometry).
  2. Wash the cells once with Hanks’ BSS + 10% serum.
  3.  

  4. Disrupt the cell pellet by “racking” the tube.
  5. Add 2 mL 1X M-Lyse Buffer per spleen processed.
  6. Incubate the cells at room temperature until lysis is complete (10 minutes).
  7.  

  8. Wash the leukocytes with 2 mL 1X Wash Buffer.
  9. Resuspend the cells in 1 mL 1X Wash Buffer.
  10.  

  11. Fix the cells with 100 uL 10X Fixative Concentrate if flow cytometry analysis will be delayed more than one hour.
  12. Or
  13. Utilize leukocytes for alternate downstream applications
  14.  

    If the cell preparation will be further processed with a T cell enrichment column, complete these steps prior to treating cells with H-Lyse Buffer.

  15. Prepare a single cell suspension of mononuclear cells.
  16. Wash the cells with excess sterile PBS.
  17.  

Technical Hints

  • If flow cytometric analysis of the cells will be delayed for more than 1 hour, the cells can be fixed at this time to stabilize the cells for later analysis. This step should be eliminated if cells are to be used for culture.
  • Cells should be stored at 2 °C to 8 °C until analysis. Although stained cells will be stable for up to 48 hours, we recommend that flow cytometric analysis be performed as soon as possible.

Reagents Provided

Citations for Mouse Erythrocyte Lysing Kit

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

21 Citations: Showing 1 - 10
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  1. CC chemokine CCL1 receptor CCR8 mediates conversion of mesenchymal stem cells to embryoid bodies expressing FOXP3+CCR8+ regulatory T cells
    Authors: NS Haque, A Tuteja, N Haque
    PLoS ONE, 2019;14(7):e0218944.  2019
  2. CC chemokine CCL1 receptor CCR8 mediates conversion of mesenchymal stem cells to embryoid bodies expressing FOXP3+CCR8+ regulatory T cells
    Authors: NS Haque, A Tuteja, N Haque
    PLoS ONE, 2019;14(7):e0218944.  2019
  3. Frontline Science: Employing enzymatic treatment options for management of ocular biofilm-based infections
    Authors: A Kugadas, J Geddes-McA, E Guy, A DiGiandome, DB Sykes, MK Mansour, R Mirchev, M Gadjeva
    J. Leukoc. Biol., 2019;0(0):.  2019
  4. Unexpected kidney-restricted role for IL-17 receptor signaling in defense against systemic Candida albicans infection
    Authors: K Ramani, CV Jawale, AH Verma, BM Coleman, JK Kolls, PS Biswas
    JCI Insight, 2018;3(9):.  2018
  5. In vivo neutralization of the protagonist role of macrophages during the chronic inflammatory stage of Huntington's disease
    Authors: J Pido-Lopez, R Andre, AC Benjamin, N Ali, S Farag, SJ Tabrizi, GP Bates
    Sci Rep, 2018;8(1):11447.  2018
  6. Protein kinase C theta is dispensable for suppression mediated by CD25+CD4+ regulatory T cells
    Authors: K Siegmund, N Thuille, K Wachowicz, N Hermann-Kl, G Baier
    PLoS ONE, 2017;12(5):e0175463.  2017
  7. A humanized mouse-based HIV-1 viral outgrowth assay with higher sensitivity than in vitro qVOA in detecting latently infected cells from individuals on ART with undetectable viral loads
    Authors: P Charlins, K Schmitt, L Remling-Mu, LE Hogan, E Hanhauser, KS Hobbs, F Hecht, SG Deeks, TJ Henrich, R Akkina
    Virology, 2017;507(0):135-139.  2017
  8. Virus-induced asthma attack: The importance of allergic inflammation in response to viral antigen in an animal model of asthma
    Authors: C Skappak, R Ilarraza, YQ Wu, MG Drake, DJ Adamko
    PLoS ONE, 2017;12(7):e0181425.  2017
  9. A humanized mouse model for HIV-2 infection and efficacy testing of a single-pill triple-drug combination anti-retroviral therapy
    Authors: Shuang Hu
    Virology, 2017;501(0):115-118.  2017
  10. Defect density in multiwalled carbon nanotubes influences ovalbumin adsorption and promotes macrophage activation and CD4(+) T-cell proliferation
    Int J Nanomedicine, 2016;11(0):4357-71.  2016
  11. Eriobotrya japonica Water Extract Characterization: An Inducer of Interferon-Gamma Production Mainly by the JAK-STAT Pathway
    Authors: Khalid Z Matalka
    Molecules, 2016;21(6):.  2016
  12. Immunogenicity without Efficacy of an Adenoviral Tuberculosis Vaccine in a Stringent Mouse Model for Immunotherapy during Treatment.
    Authors: Alyahya S, Nolan S, Smith C, Bishai W, Sadoff J, Lamichhane G
    PLoS ONE, 2015;10(5):e0127907.  2015
  13. HIV pre-exposure prophylaxis: mucosal tissue drug distribution of RT inhibitor Tenofovir and entry inhibitor Maraviroc in a humanized mouse model.
    Authors: Veselinovic M, Yang K, Lecureux J, Sykes C, Remling-Mulder L, Kashuba A, Akkina R
    Virology, 2014;464(0):253-63.  2014
  14. In vivo blockade of the PD-1 receptor suppresses HIV-1 viral loads and improves CD4+ T cell levels in humanized mice.
    Authors: Palmer B, Neff C, Lecureux J, Ehler A, Dsouza M, Remling-Mulder L, Korman A, Fontenot A, Akkina R
    J Immunol, 2013;190(1):211-9.  2013
  15. Brain human monoclonal autoantibody from sydenham chorea targets dopaminergic neurons in transgenic mice and signals dopamine D2 receptor: implications in human disease.
    Authors: Cox, Carol J, Sharma, Meenaksh, Leckman, James F, Zuccolo, Jonathan, Zuccolo, Amir, Kovoor, Abraham, Swedo, Susan E, Cunningham, Madelein
    J Immunol, 2013;191(11):5524-41.  2013
  16. Ligand Modulation of the Epstein-Barr Virus-induced Seven-transmembrane Receptor EBI2: IDENTIFICATION OF A POTENT AND EFFICACIOUS INVERSE AGONIST.
    Authors: Benned-Jensen T, Smethurst C, Holst PJ, Page KR, Sauls H, Sivertsen B, Schwartz TW, Blanchard A, Jepras R, Rosenkilde MM
    J. Biol. Chem., 2011;286(33):29292-302.  2011
  17. Identification of transforming growth factor beta1-driven genetic programs of acute lung fibrosis.
    Authors: Pulichino AM, Wang IM, Caron A, Mortimer J, Auger A, Boie Y, Elias JA, Kartono A, Xu L, Menetski J, Sayegh CE
    Am. J. Respir. Cell Mol. Biol., 2008;39(3):324-36.  2008
  18. The angiogenic response is dictated by beta3 integrin on bone marrow-derived cells.
    Authors: Feng W, McCabe NP, Mahabeleshwar GH, Somanath PR, Phillips DR, Byzova TV
    J. Cell Biol., 2008;183(6):1145-57.  2008
  19. Liposomal retinoic acids modulate asthma manifestations in mice.
    Authors: Maret M, Ruffie C, Periquet B, Campo AM, Menevret M, Phelep A, Dziewiszek K, Druilhe A, Pretolani M
    J. Nutr., 2007;137(12):2730-6.  2007
  20. Gamma interferon does not enhance clearance of Pseudomonas aeruginosa but does amplify a proinflammatory response in a murine model of postseptic immunosuppression.
    Authors: Murphey ED, Herndon DN, Sherwood ER
    Infect. Immun., 2004;72(12):6892-901.  2004
  21. Dual roles of IL-4 in lung injury and fibrosis.
    Authors: Huaux F, Liu T, McGarry B, Ullenbruch M, Phan SH
    J. Immunol., 2003;170(4):2083-92.  2003

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Reviews for Mouse Erythrocyte Lysing Kit

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Mouse Erythrocyte Lysing Kit
By Anonymous on 04/04/2019
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Mouse Erythrocyte Lysing Kit
By Anonymous on 03/21/2019
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Mouse Erythrocyte Lysing Kit
By Anonymous on 12/14/2016
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Mouse Erythrocyte Lysing Kit is used for the removal of erythrocytes (RBC) from splenocyte preparations. It can gently lyses erythrocytes, stabilizes leukocytes for analysis by flow cytometry and maintains natural light scattering and fluorescent staining properties of leukocytes. Maintains viability of leukocytes for subsequent tissue culture manipulation. We use used this kit for the removal of erythrocytes (RBC) from splenocyte preparations. This kit is part of the MagCellect CD24- CD44+ Breast Cancer Stem Cell Isolation Kit. I recommend it to be used for the removal of erythrocytes (RBC) from splenocyte preparations.