BD, 560123, BD Pharmingen™ Alexa Fluor® 555 Mouse anti-Human TRA-1-81 Antigen

Reactivity: Human (QC Testing), Rhesus (Reported)
Isotype: Mouse BALB/c IgM, κ
Immunogen: Human Embryonal Carcinoma Cell Line
Application: Bioimaging (Routinely Tested)
Vol. Per Test: 5 µl
RRID: AB_1645392
Storage Buffer: Aqueous buffered solution containing BSA, protein stabilizer, and ≤0.09% sodium azide.
Regulatory Status: RUO
Preparation And Storage: Preparation And Storage Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to Alexa Fluor® 555 under optimum conditions, and unreacted Alexa Fluor® 555 was removed.
Recommended Assay Procedures: Recommended Assay Procedures Recommended Assay Procedure: 1. Seed the cells in appropriate culture medium at an appropriate cell density in a BD Falcon™ 96-well Imaging Plate (Cat. No. 353219), and culture overnight to 48 hours. 2. Remove the culture medium from the wells, wash the wells twice with 100 μl of 1× PBS, and fix the cells by adding 100 µl of fresh 3.7% Formaldehyde in PBS or BD Cytofix™ fixation buffer (Cat. No. 554655) to each well and incubating for 10 minutes at room temperature (RT). 3. Remove the fixative from the wells, and wash the wells twice with 100 μl of 1× PBS. 4. Dilute the antibody in 1× PBS, and stain the cells by adding 50 µl of the diluted antibody conjugate to each well and incubating for 1 hour at RT. 5. Remove the diluted antibody, and wash the wells twice with 100 μl of 1× PBS. 6. Remove the PBS, and counter-stain the nuclei by adding 100 μ l of a 2 μ g/ml solution of Hoechst 33342 (eg, Sigma-Aldrich Cat. No. B2261) in 1× PBS to each well at least 15 minutes before imaging. 7. View and analyze the cells on an appropriate imaging instrument.