Mouse Erythrocyte Lysing Kit

R&D Systems | Catalog # WL2000

R&D Systems
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Key Product Details

Species

Mouse

Product Summary for Mouse Erythrocyte Lysing Kit

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. 

Formulation, Preparation, and Storage

Shipping

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.

Product Documents for Mouse Erythrocyte Lysing Kit

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Mouse Erythrocyte Lysing Kit

For research use only

⚠ 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.

Citations for Mouse Erythrocyte Lysing Kit

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  • Mouse Erythrocyte Lysing Kit
    Name: Dhivya Sridaran
    Verified Customer | Posted 04/04/2019
  • Mouse Erythrocyte Lysing Kit
    Name: Shambhavi Bhagwat
    Verified Customer | Posted 03/21/2019
  • Mouse Erythrocyte Lysing Kit
    Name: Anonymous
    Verified Customer | Posted 12/14/2016
    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.

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Protocols

View specific protocols for Mouse Erythrocyte Lysing Kit (WL2000):

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

 

 
 
 
 

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.

 
  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. Disrupt the cell pellet by “racking” the tube.
  4. Add 2 mL 1X M-Lyse Buffer per spleen processed.
  5. Incubate the cells at room temperature until lysis is complete (10 minutes).
  6. Wash the leukocytes with 2 mL 1X Wash Buffer.
  7. Resuspend the cells in 1 mL 1X Wash Buffer.
  8. Fix the cells with 100 uL 10X Fixative Concentrate if flow cytometry analysis will be delayed more than one hour.
  9. Or
  10. Utilize leukocytes for alternate downstream applications
  11. Prepare a single cell suspension of mononuclear cells.
  12. Wash the cells with excess sterile PBS.

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.

FAQs

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