IEL Shake Medium

Dillon Corvino

EDTA/RPMI medium used for the mechanical shake washes after DTT/EDTA epithelial stripping.
Published

September 18, 2026

Protocol ID: BUF-IEL-SHAKE Version: v1.0 Author: Dillon Corvino Date: September 18, 2026

Purpose

Used for the manual shake wash steps that mechanically dislodge residual intraepithelial lymphocytes (IELs) from intestinal tissue after DTT/EDTA stripping.

Linked protocols

  • MUS-IEL-ISO

Working buffer composition

25 mL per mouse (2 x 10 mL for shake rounds + ~5 mL margin).

EDTA stock: 500 mM. For 2 mM final in 25 mL: add 100 uL EDTA stock.

Component Final concentration Volume for 25 mL
RPMI 1640 Base 23.6 mL
L-glutamine (200 mM stock) 1% (v/v) 250 uL
Penicillin/Streptomycin (100x stock) 1% (v/v) 250 uL
HEPES (1 M stock) 2% (v/v) 500 uL
EDTA (500 mM stock) 2 mM 100 uL

Preparation

A. Preparation of stock (if applicable)

Not applicable — prepared directly as the working buffer.

B. Preparation of working buffer

  1. Combine RPMI 1640 with L-glutamine, Penicillin/Streptomycin, HEPES, and EDTA per the table above.
  2. Mix gently by inversion.

Storage and stability

  • Can be prepared the evening before use and held at 4 °C.

Reagent details

Component Supplier Cat# Notes
RPMI 1640 [TBD] [TBD]
L-glutamine (200 mM stock) [TBD] [TBD]
Penicillin/Streptomycin (100x) [TBD] [TBD]
HEPES (1 M stock) [TBD] [TBD]
EDTA (0.5 M stock) [TBD] [TBD]

Safety (brief)

Not applicable — no reagent-specific hazards beyond standard laboratory PPE.

Version history

Version Date Author Change summary
v1.0 2026-09-18 Dillon Corvino Split out of prot_mus_iel_isolation.qmd into its own buffer document (buffer/protocol architecture redesign) — no change to composition.