Immunocytochemistry/ Immunofluorescence abreview for Anti-SCF antibody [EP665Y] (Alexa Fluor® 594)

Immunocytochemistry/ Immunofluorescence

Human Cell (HL-60 cell line)

No

HL-60 cell line

BSA 5%, FBS 10% as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C

Formaldehyde

1 hour(s) and 30 minute(s) · Temperature: 21°C

1/50

None used

HL-60 cells were seeded in plates that contained cover slips and were pre-coated with L-ornithine. Incubation lasted for 2 hours in 37oC. After that, the supernatant was removed and cells was washed with PBS 1x (3 times). Formaldehyde 3,7% was used for fixation (incubation: 10 minutes). Supernatant was again removed and blocking step followed (BSA 5%, FBS 10%, 2 hours, RT). During the probing stage, cover slips were carefully removed and 100 ul of ab215092 (anti-SCF)/ab208568 (isotypic control) was used upon them (dilution 1:50 for both antibodies in the blocking medium). The probing stage lasted for 1,5 hour at RT. In the end, fluorescence was detected under a fluorescence microscope under the proper channel (Texas Red).