Frozen samples of total epididymal cells from 10-day old rats were thawed on ice and gently resuspended in DMEM medium containing 10% FBS. Cells were centrifuged (300g; 7 min). The supernatant was discarded and the pelleted cells were suspended in FACS buffer for counting (0.1% BSA in 1X PBS). Cells were distributed as 700 000 cell aliquots into 1.5 ml tubes, centrifuged (300g; 7 min) and resuspended in blocking buffer (FACS buffer containing 2% BSA and 10% normal donkey serum). Cells were incubated on ice for 30 min. A mouse monoclonal anti-rat CD49f antibody conjugated with Alexa Fluor® 647 (CD49F; 0.5µg/µl; Bio-Rad Laboratories, Mississauga, ON) was then added to the cells, and the mixture was incubated on ice for 1h in the dark. Cells were washed two times with 1 ml of FACS buffer and resuspended in 350 ul of FACS buffer containing Hoechst dye (0.05 µg/ml; Biotium) and incubated on ice for 10 mins. Cells were then washed once with 1 ml of FACS buffer, resuspended in 350 ul of FACS buffer and transferred to FACS tubes. Flow cytometric analyses were done using an LSR Fortessa and Cell Quest Pro software (BD Biosciences, Franklin Lakes, NJ).
Alexa fluor 647 cd49f
Manufactured by Bio-Rad
Alexa Fluor® 647 (CD49F) is a fluorescent dye used for labeling and detection in flow cytometry and other fluorescence-based applications. It is a covalently linked conjugate of the Alexa Fluor® 647 dye and an antibody specific for the CD49F cell surface marker.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using alexa fluor 647 cd49f
1
Rat Epididymal Cell Isolation and Flow Cytometry
2
Rat Epididymal Cell Isolation and Flow Cytometry
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Frozen samples of total epididymal cells from 10-day old rats were thawed on ice and gently resuspended in DMEM medium containing 10% FBS. Cells were centrifuged (300g; 7 min). The supernatant was discarded and the pelleted cells were suspended in FACS buffer for counting (0.1% BSA in 1X PBS). Cells were distributed as 700 000 cell aliquots into 1.5 ml tubes, centrifuged (300g; 7 min) and resuspended in blocking buffer (FACS buffer containing 2% BSA and 10% normal donkey serum). Cells were incubated on ice for 30 min. A mouse monoclonal anti-rat CD49f antibody conjugated with Alexa Fluor® 647 (CD49F; 0.5µg/µl; Bio-Rad Laboratories, Mississauga, ON) was then added to the cells, and the mixture was incubated on ice for 1h in the dark. Cells were washed two times with 1 ml of FACS buffer and resuspended in 350 ul of FACS buffer containing Hoechst dye (0.05 µg/ml; Biotium) and incubated on ice for 10 mins. Cells were then washed once with 1 ml of FACS buffer, resuspended in 350 ul of FACS buffer and transferred to FACS tubes. Flow cytometric analyses were done using an LSR Fortessa and Cell Quest Pro software (BD Biosciences, Franklin Lakes, NJ).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!