PBE buffer: Prepare a solution containing phosphate-buffered saline (PBS), pH 7.2, 0.5 % bovine serum albumin (BSA), and 2  mM EDTA by diluting MACS BSA Stock Solution 1:20 with autoMACS Rinsing Solution. Keep buffer cold (2−8 °C). Prepare buffer fresh and degas prior to use.
▲ Note: EDTA can be replaced by other supplements such as anticoagulant citrate dextrose formula-A (ACD-A) or citrate phosphate dextrose (CPD). BSA can be replaced by other proteins such as mouse serum albumin, mouse serum, or fetal bovine serum (FBS). Buffers or media containing Ca²⁺ or Mg²⁺ are not recommended for use.

Create a viable single-cell suspension from a solid mouse tumor using the gentleMACS™ Octo Dissociator with Heaters in combination with the Tumor Dissociation Kit, mouse. Follow the protocol of the kit data sheet.

Download data sheet

Tumor Dissociation Kit, mouse
 

Enrich tumor-infiltrating leukocytes using CD45 (TIL) MicroBeads, mouse, and LS or MS Columns. Follow the protocol of the kit data sheet.

We recommend filtering the magnetically labeled cell suspension to guarantee it is single-celled before separating it on the column.

1. Place a Pre-Separation Filter (30 µm) on the LS or MS Column.
2. Rinse the column 3 times with PBE buffer, ensuring that the filter is pre-wetted.
3. Apply the cell suspension and washing buffer to the filter on the column.

Download the data sheet

CD45 (TIL) MicroBeads, mouse

The following is a list of additional resources that may be of interest:
 

• Preservation of cell surface epitopes
  
• Convenient tumor dissociation
• REAfinity™ Antibodies – Flow cytometry is in their genes
• REAfinity Antibodies – Frequently asked questions
• Brauner, J. et al. (2017) Workflow automation and parallelization improves the isolation and analysis of tumor-infiltrating immune cell populations. AACR, Washington D.C., USA.
• Miltenyi Biotec product list: Cancer antibodies, mouse
• Application Note: Functional TILs from human renal cell carcinoma. Baldan, V. et al., Manchester Academic Healthcare Science Centre.

Unless otherwise specifically indicated, Miltenyi Biotec products and services are for research use only and not for therapeutic or diagnostic use.

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