LentiBrite Histone H2B-RFP Lentivi

LentiBrite Histone H2B-RFP Lentivi

Stock Code: 3586362
Manufacturer Part No: 17-10228
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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-RFP signal shows interphase nuclei, with the lower nuclei connected by a strand representing a segregation defect.

Additional cell types and Hard-to-transfect cell types:
(See Figure 2 in datasheet)
Primary HUVECs were transduced at an MOI of 20. The image depicts an irregular interphase nucleus. HT1080 were transduced at an MOI of 40 for 24 hours. The image depicts an interphase nucleus. Human mesenchymal stem cells (HuMSC) transduced with lentiviral particles at an MOI of 40. The cell is in anaphase and displays lagging chromosomes.

Mitosis in Various Stages:
(See Figure 3 in datasheet)
U2OS cells 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-TagRFP 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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