. Author manuscript; available in PMC: 2012 Jun 25.

Published in final edited form as: Nat Protoc. 2010 Dec 9;6(1):1–14. doi: 10.1038/nprot.2010.168

Table 1.

Antibodies and fluorophores for HSPC harvesting.

Antigen	Conjugate	Clone	Vendor	Cat. no.	Dilution
Lineage cocktail
Gr1	Biotin	RB6-8C5	BD Biosciences	553125	—^a
Mac1	Biotin	M1/70	BD Biosciences	553309	—^a
B220	Biotin	Ra3-6B2	BD Biosciences	553086	—^a
Ter119	Biotin	Ter119	BD Biosciences	553672	—^a
CD4	Biotin	GK1.5	BD Biosciences	553728	—^a
CD8	Biotin	53-6.7	BD Biosciences	553029	—^a
CD3	Biotin	145-2C11	BD Biosciences	553060	—^a
FACS
c-Kit	APC	2B8	BioLegend	105812	1:100
Sca-1	Pacific blue (PB)	D7	BioLegend	108120	1:100
CD34	FITC	RAM34	eBioscience	11-0341-85	1:100
Flk2	Phycoerythrin (PE)	A2F10.1	BD Biosciences	553542	1:200
CD48	FITC	HM48-1	BioLegend	103404	1:1000
CD150	PE	Tc15-12F12.2	BioLegend	115904	1:100
Streptavidin	Pacific orange (PO)		Invitrogen	S32365	1:500

A selected combination of these markers and fluorophores can be used for HSPC sorting to obtain LKS CD34 ⁻Flk2⁻ or LKS CD150⁺CD48⁻ cell populations. The ideal color assortment and supplier can vary depending on the cell sorter used. Note that streptavidin staining allows gating out the remaining lineage-positive cells during the sorting.

For dilution, see REAGENT SETUP and Step 5 of PROCEDURE.