LentiBrite Histone H2B-GFP Lentivi

LentiBrite Histone H2B-GFP Lentivi

Stock Code: 3586363
Manufacturer Part No: 17-10229
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£998.86 (exc VAT) per EACH
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Application


Research Sub CategoryApoptosis - AdditionalCell Cycle, DNA Replication & Repair


Fluorescence microscopy imaging:
(See Figure 1 in datasheet)
HeLa cells were plated in a chamber slide and transduced with lentiviral particles at an MOI of 20 for 24 hours. After media replacement and 48 hours further incubation, cells were fixed with formaldehyde and mounted. Images were obtained by oil immersion wide-field fluorescence microscopy. The histone H2B-GFP appears solely associated with mitotic chromosomes.

Additional cell types and Hard-to-transfect cell types:
(See Figure 2 in datasheet)
U2OS cells transduced at an MOI of 20. The top cell displays telophase chromosomes, and the bottom cell contains anaphase chromosomes. HT1080 were transduced at an MOI of 40 for 24 hours. The two nuclei are in interphase. Primary cell types Human mesenchymal stem cells (HuMSC) were plated in chamber slides and transduced with lentiviral particles at an MOI of 40 for 24 hours. The three nuclei are in interphase.

Difficult-to-transfect cell types at various mitosis stages:
(See Figure 3 in datasheet)
Primary cell types HUVEC were plated in chamber slides and transduced with lentiviral particles at an MOI of 20 for 24 hours. The images show cells in various stages of mitosis.

For optimal fluorescent visualization, it is recommended to analyze the target expression level within 24-48 hrs after transfection/infection for optimal live cell analysis, as fluorescent intensity may dim over time, especially in difficult-to-transfect cell lines. Infected cells may be frozen down after successful transfection/infection and thawed in culture to retain positive fluorescent expression beyond 24-48 hrs. Length and intensity of fluorescent expression varies between cell lines. Higher MOIs may be required for difficult-to-transfect cell lines.


Research CategoryApoptosis & CancerEpigenetics & Nuclear Function


Components


Histone H2B-TagGFP2 Lentivirus: One vial containing 25 µL of lentiviral particles at a minimum of 3 x 10E8 infectious units (IFU) per mL. For lot-specific titer information, please see lot specific “Viral Titer” in the product specifications of the datasheet.PromoterEF-1 (Elongation Factor-1)Multiplicty of Infection (MOI)MOI = Ratio of # of infectious lentiviral particles (IFU) to # of cells being infected.Typical MOI values for high transduction efficiency and signal intensity are in the range of 20-40. For this target, some cell types may require lower MOIs (e.g., HT-1080, HeLa, human mesenchymal stem cells (HuMSC)), while others may require higher MOIs (e.g., human umbilical vein endothelial cells (HUVEC), U2OS). NOTE: MOI should be titrated and optimized by the end user for each cell type and lentiviral target to achieve desired transduction efficiency and signal intensity.


General description


Chromatin, the higher order structure of DNA and nucleosomes, constitutes the majority of the nucleus of the eukaryotic cell. Changes in chromatin structure are the essence of many essential nuclear processes, including transcription, mitosis, meiosis, and apoptosis. The nucleosome comprises an octomer of four core histone proteins (H2A, H2B, H3 and H4)

Quality Level100
ManufacturerSIGMA-ALDRICH
Technique(s)immunocytochemistry: suitable, cell based assay: suitable, transfection: suitable, immunofluorescence: suitable

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