After 72 hours of suspension culture with drug exposure, CD34+ CML cells were stained with p-CRKL (Cell Signaling) or p-STAT5 antibodies (Cell Signaling) overnight, followed by incubation with a secondary antibody (anti–rabbit IgG FITC-conjugate, Invitrogen) prior to FACS analysis as previously described [53 (link)]. p-CRKL and p-STAT5 levels were determined as the geometric mean fluorescence intensity (MFI) subtracted by the MFI of cells stained with isotype IgG control, and were normalized as a percentage of control cells incubated for the same time with DMSO only.
Anti rabbit igg fitc conjugate
Manufactured by Thermo Fisher Scientific
Anti–rabbit IgG FITC-conjugate is a secondary antibody that binds to rabbit immunoglobulin G (IgG) and is conjugated to the fluorescent dye FITC (fluorescein isothiocyanate). It is used in various laboratory techniques, such as immunofluorescence, Western blotting, and flow cytometry, to detect and visualize rabbit primary antibodies.
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2 protocols using anti rabbit igg fitc conjugate
1
Quantifying CML Cell Signaling
2
Immunofluorescence Assay for SIRT1 Localization
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Cells were seeded on glass bottom dishes (Iwaki, Asahi Techno Glass) and fixed with 4% paraformaldehyde for 15 minutes and blocked with 1% BSA, 0.1% Triton X-100 in PBS for 20 min. Cells were then incubated with SIRT1 antibody (1∶200; Abcam Ab13749) in blocking buffer for 1 hour. After 3 PBS washes, cells were incubated with anti-rabbit IgG FITC conjugate (1∶500; Invitrogen cat# A11008) for 30 minutes under light protection. After 3 PBS washes, cells were imaged using a LSM 710 confocal microscope (Zeiss). AQM Advance 6.0 software (Kinetic Imaging) was used for image analysis.
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