• MACSmix™ Tube Rotator (# 130-090-753) in combination with an incubator at 37 °C
• gentleMACS™ Dissociator (# 130-093-235), gentleMACS Octo Dissociator (# 130-095-937), or gentleMACS Octo Dissociator with Heaters (# 130-096-427)
• gentleMACS C Tubes (# 130-093-237, # 130-096-334)
• (Optional) ART® 1000 REACH™ pipet tips (Molecular BioProducts, Inc.) for removal of dissociated material from the closed C Tubes.

PBE buffer for spleen/lung tissue dissociation and for cell isolation

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 (# 130-091-376) 1:20 with autoMACS® Rinsing Solution (# 130-091-222). Keep buffer cold (2–8 °C). 
▲ Note: Always use freshly prepared buffer. Do not use autoMACS Running Buffer or MACSQuant® Running Buffer as they contain a small amount of sodium azide that could affect the results.

• Spleen Dissociation Kit, mouse (# 130-095-926)
• Phosphate-buffered saline pH 7.2
  
• Pre-Separation Filters, 70 μm (# 130-095-823)
  

• Lung Dissociation Kit, mouse (# 130-095-927)
• Phosphate-buffered saline pH 7.2
  
• MACS SmartStrainers (70 µm) (# 130-098-462)
  

• Lamina Propria Dissociation Kit, mouse (# 130-097-410)
• MACS SmartStrainers, 100 μm (# 130-098-463)
• (Sterile) 1× Hank’s balanced salt solution (HBSS) without Ca²⁺ and Mg²⁺ containing 10 mM HEPES. Store at room temperature. This solution is referred to as HBSS (w/o) in the protocol of the kit data sheet.
• (Sterile) 1× HBSS with Ca²⁺ and Mg²⁺ containing 10 mM HEPES. Store at room temperature. This solution is referred to as HBSS (w) in the protocol of the kit data sheet.
• (Sterile) Predigestion solution: prepare 1× HBSS (w/o) containing 5 mM EDTA, 5% fetal bovine serum (FBS), 1 mM DTT freshly before each digestion. Store at room temperature.
  ▲ Note: Per digestion a volume of 40 mL of the predigestion solution is required. 
• (Sterile) Digestion solution: prepare 1× HBSS (w) containing 5% FBS freshly before each digestion. Store at room temperature.
  ▲ Note: Per digestion a volume of 2.5 mL of the digestion solution is required. 
• (Sterile) PB buffer: prepare a solution containing phosphate-buffered saline (PBS), pH 7.2, and 0.5 % bovine serum albumin (BSA). Keep buffer cold (2–8 °C).
  ▲ Note: BSA can be replaced by other proteins such as mouse serum albumin, mouse serum, or FBS. 

• MACSQuant Analyzer 10 (# 130-096-343)
  
• MACS Columns and MACS Separators: CD11c⁺ cells can be enriched using MS, LS, or XS Columns. Positive selection can also be performed on the autoMACS Pro Separator.

▲Note: Column adapters are required to insert certain columns into the VarioMACS™ or SuperMACS™ II Separators. For details refer to the respective MACS Separator data sheet. 

• (Optional) Fluorochrome-conjugated CD11c antibodies for flow cytometry analysis, e.g., CD11c-VioBlue® (# 130‑102‑413). Learn more about our antibodies and dyes.
• (Optional) FcR Blocking Reagent, mouse (# 130-092-575) to avoid Fc receptor–mediated antibody labeling 
• (Optional) Propidium Iodide Solution (# 130-093-233) or 7-AAD for flow cytometry exclusion of dead cells
• (Optional) Dead Cell Removal Kit (# 130-090-101) for the depletion of dead cells
• (Optional) Pre-Separation Filters, 70 μm (# 130-095-823) to remove cell clumps