Fixed and blocked HeLa CCL-2 cells were incubated with 10 μg/mL cleavable biotin-labeled rabbit anti-Ki67 (Thermo Fisher Scientific) for 45 min. Subsequently, cells were stained by 10 μg/mL cleavable fluorescent streptavidin. Following that, one, two, three and four rounds of amplification were applied to different sets of cells. In each round of amplification, the cells were first incubated with cleavable biotin-labeled goat-anti-chicken antibodies and then with cleavable fluorescent streptavidin. Biotin and fluorophores were cleaved by TCEP (100 mM, pH = 9.5). The cells were blocked with 0.5 mg/mL biotin. The cells were incubated with cleavable biotin-labeled goat anti-chicken and then cleavable fluorescent streptavidin.
Cleavable biotin labeled rabbit anti ki67
Manufactured by Thermo Fisher Scientific
Cleavable biotin-labeled rabbit anti-Ki67 is a laboratory reagent designed for the detection and quantification of the Ki-67 protein, a well-established marker of cellular proliferation. This product features a biotin label, which can be cleaved off, allowing for further downstream applications.
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3 protocols using cleavable biotin labeled rabbit anti ki67
1
Amplified Protein Visualization in HeLa Cells
2
Amplified Ki67 Detection in HeLa Cells
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Fixed and blocked HeLa CCL-2 cells were incubated with 10 μg/mL cleavable biotin-labeled rabbit anti-Ki67 (Thermo Fisher Scientific) for 45 min. Subsequently, the cells were stained by 10 μg/mL cleavable fluorescent streptavidin. Then, one, two, three and four rounds of amplification were applied to different sets of cells. In each round of amplification, the cells were first incubated with cleavable biotin-labeled goat-anti-chicken antibodies and then with cleavable fluorescent streptavidin. The cells were then incubated with TCEP (100 mM, pH = 9.5) for 30 min at 37 °C. Subsequently, the cells were washed three times with PBT and three times with 1X PBS, each for 5 min.
3
Amplified Biotin-Streptavidin Imaging
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Fixed and blocked HeLa CCL-2 cells were incubated with 10 μg/mL cleavable biotin-labeled rabbit anti-Ki67 (Thermo Fisher Scientific) for 45 min. Subsequently, cells were stained by 10 μg/mL cleavable fluorescent streptavidin. Following that, one, two, three and four rounds of amplification were applied to different sets of cells. In each round of amplification, the cells were first incubated with cleavable biotin-labeled goat-anti-chicken antibodies and then with cleavable fluorescent streptavidin. Biotin and fluorophores were cleaved by TCEP (100 mM, pH = 9.5). The cells were then incubated with cleavable fluorescent streptavidin.
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