Anti-Integrin β1 Antibody, clone MB1.2

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	The cells were trypsinized and washed twice with 0.5% BSA in PBS. The cells were washed twice and incubated with allophycocyanin-conjugated anti-rat or anti-mouse secondary antibody for an additional 30 min on ice.	After further washing, flow cytometric analysis was performed on a FACSCanto flow cytometer (BD Biosciences), and flow data overlay plots were produced using CellQuest Pro software (BD Biosciences).

Protocol tips
The cells were trypsinized and washed twice with 0.5% BSA in PBS. The cells were washed twice and incubated with allophycocyanin-conjugated anti-rat or anti-mouse secondary antibody for an additional 30 min on ice.
Downstream tips
After further washing, flow cytometric analysis was performed on a FACSCanto flow cytometer (BD Biosciences), and flow data overlay plots were produced using CellQuest Pro software (BD Biosciences).

Publication protocol

4.1G+/+ and 4.1G−/− MEF cells were serum-starved for 18 h. The cells were trypsinized and washed twice with 0.5% BSA in PBS. Primary antibodies against total β1 integrin (catalog no. MAB1997, Millipore) and against active-form β1 integrin (clone 9EG7, BD Biosciences) were used to stain the cells in 0.5% BSA in PBS for 30 min on ice. The cells were washed twice and incubated with allophycocyanin-conjugated anti-rat or anti-mouse secondary antibody for an additional 30 min on ice. After further washing, flow cytometric analysis was performed on a FACSCanto flow cytometer (BD Biosciences), and flow data overlay plots were produced using CellQuest Pro software (BD Biosciences).

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