3% Medium (IEL Wash/Base Medium)
Dillon Corvino
Low-serum RPMI wash medium used for intestinal washes and as the base for IEL Strip Medium.
Protocol ID: BUF-IEL-3PCT Version: v1.0 Author: Dillon Corvino Date: September 18, 2026
Purpose
Low-serum RPMI wash medium used to wash the opened small intestine before stripping, and as the base medium for IEL Strip Medium (BUF-IEL-STRIP).
Linked protocols
MUS-IEL-ISO
Working buffer composition
50 mL per mouse (20 mL for intestinal washes + 20 mL as base for Strip Medium + ~10 mL margin).
| Component | Final concentration | Volume for 50 mL |
|---|---|---|
| RPMI 1640 | Base | 46.0 mL |
| FCS | 3% (v/v) | 1.5 mL |
| HEPES (1 M stock) | 2% (v/v) | 1.0 mL |
| Penicillin/Streptomycin (100x stock) | 1% (v/v) | 0.5 mL |
| L-glutamine (200 mM stock) | 1% (v/v) | 0.5 mL |
Preparation
A. Preparation of stock (if applicable)
Not applicable — prepared directly as the working buffer.
B. Preparation of working buffer
- Combine RPMI 1640 with FCS, HEPES, Penicillin/Streptomycin, and L-glutamine per the table above.
- Mix gently by inversion.
Storage and stability
- Can be prepared the evening before use and held at 4 °C.
- Bring to room temperature or 37 °C (as needed) before use.
Reagent details
| Component | Supplier | Cat# | Notes |
|---|---|---|---|
| RPMI 1640 | [TBD] | [TBD] | |
| FCS | [TBD] | [TBD] | |
| HEPES (1 M stock) | [TBD] | [TBD] | |
| Penicillin/Streptomycin (100x) | [TBD] | [TBD] | |
| L-glutamine (200 mM stock) | [TBD] | [TBD] |
Safety (brief)
- FCS is of animal origin and should be treated as a potential biohazard.
- Not applicable 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. |