Jul 23, 2020
SOLUTION- 06 - Lysis Buffer
- Marco Cosentino1,
- Elisa Storelli1,
- Alessandra Luini1,
- Massimiliano LM Legnaro1,
- Emanuela Rasini1,
- Marco Ferrari1,
- Franca Marino1
- 1Center for Research in Medical Pharmacology, University of Insubria (Varese, Italy)
Document Citation: Marco Cosentino, Elisa Storelli, Alessandra Luini, Massimiliano LM Legnaro, Emanuela Rasini, Marco Ferrari, Franca Marino 2020. SOLUTION- 06 - Lysis Buffer. protocols.io https://dx.doi.org/10.17504/protocols.io.biwxkffn
License: This is an open access document distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Created: July 23, 2020
Last Modified: November 21, 2020
Document Integer ID: 39607
Abstract
This recipe is used in the following protocols:
- PBMC- 01a - Isolation of Human PBMC from Buffy Coat
- PBMC- 01b - Isolation of Human PBMC from Whole Blood
- PBMC- 02 - CD4+ T cell Isolation from PBMC with “Dynabeads CD4 Positive Isolation Kit”
- PBMC- 03 - TEFF+TREG Isolation from PBMC with “Miltenyi CD4+CD25+ Regulatory T cell Isolation Kit”
- PMN- 01a - Isolation of Human PMN from Buffy Coat
- PMN- 01b - Isolation of Human PMN from Whole Blood
Lysis Buffer composition g/l (M)
NH4Cl 8.248 (0.154)
KHCO3 1.0 (0.01)
EDTA 0.0368 (0.0001)
Add ultrapure water to 1 liter.
Storage: 4 °C Fridge 1 - (Room TSO8).
Filtered Lysis Buffer: filter lysis buffer using sterile syringe and filter.
Sterile filter and syringe are in closet 1 (Room PSO3).
Prymo siringe filter: catalog number EPSPE2230 Euroclone, Italy.
Storage: 4 °C Fridge 2 - (Room PSO3).
NH4Cl: catalog number 1.01145.1000 Merck, Italy.
KHCO3: catalog number 1.04854.500 Merck, Italy.
EDTA: catalog number ED2SS Sigma, Italy.