Table 1. Antibodies used in the study .... | Rockefeller University Press

Table 1.

Antibodies used in the study


Antibodies used for lineage depletion
Antigen	Dilution	Tag	Company	Clone
Anti-Mouse CD5	1:100	Biotin	eBioscience	53-7.3
Anti-Mouse CD3e	1:100	Biotin	eBioscience	145-2C11
Anti-Mouse TER-119	1:100	Biotin	eBioscience	TER-119
Anti-Mouse CD8a	1:100	Biotin	eBioscience	53-6.7
Anti-Mo/Hu CD45R (B220)	1:100	Biotin	eBioscience	RA3-6B2
Anti-Mouse Ly-6G (Gr-1)	1:100	Biotin	eBioscience	RB6-8C5
Anti-Mouse CD2	1:100	Biotin	eBioscience	RM2-5
Anti-Mo/Hu CD11b	1:100	Biotin	BioLegend	M1/70
Antibodies used for HSC sorting
Antigen	Dilution	Fluorochrome	Company	Clone
Streptavidin	1:100	PerCP	BD Pharmigen	554064
Anti-Mouse Ly-6A/E (Sca-1)	1:100	BV 510	BioLegend	D7
Anti-Mouse CD150 (signaling lymphocytic activation molecule)	1:100	BV 5605	BioLegend	TC15-12F12.2
Anti-Mouse CD117 (C-Kit)	1:100	APC-eFluor 780	Invitrogen	2B8
Anti-Mouse CD48	1:100	PE/Cy7	BioLegend	HM48-1
Primary antibodies used for cultured HSC analysis
Antigen	Dilution	Fluorochrome	Company	Clone
Anti-Mouse Ly-6A/E (Sca-1)	1:100	FITC	BioLegend	D7
Anti-Mouse EPCR	1:100	PE	BioLegend	RCR-40
Primary antibodies used for immunostaining
Host	Antibody	Dilution	Company	Cat. No.
Goat	NUMB	1:1,600	Abcam	Ab4147
Mouse	pERK (T202/Y204)	1:400	Cell Signalling	9106S
Rabbit	ERK	1:400	Cell Signalling	9102
Rabbit	pS6	1:1,200	Cell Signalling	5364S
Mouse	S6	1.400	Cell Signalling	2317S
Rabbit	mTOR S2448	1:200	Cell Signalling	5536S
Mouse	mTOR	1:200	Proteintech	66888-1
Rat	Tubulin	1:1,200	ORIGENE	SM568P
Rabbit	CDC42	1:100	Merck Millipore	07–1,466
Secondary antibodies used for immunostaining
Host	Antigen	Dilution	Company	Cat. No.	Fluorochrome
Donkey	Anti-Goat	1:800	Invitrogen	A11055	Alexa Fluor 488
Donkey	Anti-Rat	1:800	Invitrogen	A21208	Alexa Fluor 488
Donkey	Anti-Rabbit	1:800	Invitrogen	A10040	Alexa Fluor 546
Donkey	Anti-Mouse	1:800	Invitrogen	A31571	Alexa Fluor 647


Antibodies used for lineage depletion
Antigen	Dilution	Tag	Company	Clone
Anti-Mouse CD5	1:100	Biotin	eBioscience	53-7.3
Anti-Mouse CD3e	1:100	Biotin	eBioscience	145-2C11
Anti-Mouse TER-119	1:100	Biotin	eBioscience	TER-119
Anti-Mouse CD8a	1:100	Biotin	eBioscience	53-6.7
Anti-Mo/Hu CD45R (B220)	1:100	Biotin	eBioscience	RA3-6B2
Anti-Mouse Ly-6G (Gr-1)	1:100	Biotin	eBioscience	RB6-8C5
Anti-Mouse CD2	1:100	Biotin	eBioscience	RM2-5
Anti-Mo/Hu CD11b	1:100	Biotin	BioLegend	M1/70
Antibodies used for HSC sorting
Antigen	Dilution	Fluorochrome	Company	Clone
Streptavidin	1:100	PerCP	BD Pharmigen	554064
Anti-Mouse Ly-6A/E (Sca-1)	1:100	BV 510	BioLegend	D7
Anti-Mouse CD150 (signaling lymphocytic activation molecule)	1:100	BV 5605	BioLegend	TC15-12F12.2
Anti-Mouse CD117 (C-Kit)	1:100	APC-eFluor 780	Invitrogen	2B8
Anti-Mouse CD48	1:100	PE/Cy7	BioLegend	HM48-1
Primary antibodies used for cultured HSC analysis
Antigen	Dilution	Fluorochrome	Company	Clone
Anti-Mouse Ly-6A/E (Sca-1)	1:100	FITC	BioLegend	D7
Anti-Mouse EPCR	1:100	PE	BioLegend	RCR-40
Primary antibodies used for immunostaining
Host	Antibody	Dilution	Company	Cat. No.
Goat	NUMB	1:1,600	Abcam	Ab4147
Mouse	pERK (T202/Y204)	1:400	Cell Signalling	9106S
Rabbit	ERK	1:400	Cell Signalling	9102
Rabbit	pS6	1:1,200	Cell Signalling	5364S
Mouse	S6	1.400	Cell Signalling	2317S
Rabbit	mTOR S2448	1:200	Cell Signalling	5536S
Mouse	mTOR	1:200	Proteintech	66888-1
Rat	Tubulin	1:1,200	ORIGENE	SM568P
Rabbit	CDC42	1:100	Merck Millipore	07–1,466
Secondary antibodies used for immunostaining
Host	Antigen	Dilution	Company	Cat. No.	Fluorochrome
Donkey	Anti-Goat	1:800	Invitrogen	A11055	Alexa Fluor 488
Donkey	Anti-Rat	1:800	Invitrogen	A21208	Alexa Fluor 488
Donkey	Anti-Rabbit	1:800	Invitrogen	A10040	Alexa Fluor 546
Donkey	Anti-Mouse	1:800	Invitrogen	A31571	Alexa Fluor 647

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