Cells were washed once with PBS and subsequently incubated with 500 μl of nonenzymatic cell dissociation solution (Sigma) until they detached (20 min). Cells were collected in 4 ml PBS and washed twice with 1 ml of 0.1% fetal calf serum in PBS centrifuging at 500g for 5 min in between. Cells were then incubated with IIH6C4 antibody (1:200 dilution; Merck Millipore) for 40 min, with biotin-labeled MAL II (10 μg/ml; Vector Laboratories) for 40 min or with FITC–labeled PNA (10 μg/ml; Sigma–Aldrich) for 90 min. Subsequently, cells were washed twice as described previously and incubated for 20 min with 1:200 diluted goat antimouse IgM-CFL (Santa Cruz) for the cells incubated with IIH6C4 antibody or with 5 μg/ml of FITC-labeled streptavidin (Vector Laboratories) for the cells incubated with MAL II. All cells were washed twice as described previously, resuspended in 400 μl PBS, and analyzed using a FACSverse flow cytometer (BD Biosciences).
Fitc labeled pna
Manufactured by Merck Group
FITC–labeled PNA is a fluorescently labeled lectin derived from the peanut plant (Arachis hypogaea). It is used for detection and labeling of glycoconjugates.
Automatically generated - may contain errors
Lab products found in correlation
Facsverse flow cytometer, by BD (1 mentions)
Fitc labeled streptavidin, by Vector Laboratories (1 mentions)
Non enzymatic cell dissociation solution, by Merck Group (1 mentions)
Fluoromount aqueous mounting medium, by Merck Group (1 mentions)
Hoechst 33258, by Merck Group (1 mentions)
Eclipse ti microscope, by Nikon (1 mentions)
2 protocols using fitc labeled pna
1
Flow Cytometric Characterization of Cells
2
Immunofluorescent Staining of CRC Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
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MC38 and control HT-29 CRC cells were cultured in 48 well plates overnight. The cells were fixated for 15 min using 4% paraformaldehyde, blocked with PBS supplemented with 10% BSA, 10% FBS, and 0.5% Triton X-100 for 2 h at room temperature and stained overnight at 4°C with FITC-labeled PNA (50 μg/ml in PBS, Sigma-Aldrich). The cells were then incubated with Hoechst 33258 (Sigma-Aldrich) diluted 1:5,000 for 15 min at room temperature washed three times with PBS followed by mounting using Fluoromount aqueous mounting medium (Sigma-Aldrich). Pictures were obtained using the Nikon Eclipse Ti microscope.
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