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HeLa H2B-2FP

HeLa H2B-2FP

Catalogue No.

21102108

Cell Line Name

HeLa H2B-2FP

Cell Line Description

HeLa cells dually transduced with Retrovirus to stably express H2B tagged with GFP, from pWZL backbone, and H2B tagged with mCherry. Both fluorophores Sorted for low expression.

 

This cell line is distributed by ECACC on behalf of CellBank Australia, Children's Medical Research Institute, Westmead, NSW 2145, Australia. All stock supplied is manufactured and Quality Control released at source by Cell Bank Australia. Cell Bank Australia reference number:CBA-1861

Characteristics

Tissue of Origin

Cervix

Morphology

Epithelial like

Disease

Carcinoma

Culture Conditions

Cell Type

Cervical carcinoma

Subculture Routine

Remove protective cryoflex layer around the ampoule prior to thawing.


Thaw the ampoule by gently agitating in a 37oC waterbath; thawing should be rapid (around 2 minutes).


A centrifugation step to remove the cryoprotectant after thawing is necessary for this cell line.


Subculturing Procedure:


Medium Renewal: 2-3 times per week.


Subcultivation ratio: 1:8-1:16 Seeding density 0.8-1.0 x104 cells/cm2


Split subconfluent cultures (70-80%). Harvest the cells using 0.05% Trypsin/EDTA at 37°C for 5 minutes.


Culture conditions: Incubate the culture at 37oC with 10% CO2.


Cryoprotectant Medium: 10% DMSO + 90% FCS

Growth Mode

Adherent

Additional Info

Depositor

Tony Cesare - Children’s Medical Research Institute, Westmead, Australia

Country of Origin

Australia

GMO Status

Genetically Modified Organism Class 1 (GMO1)

Genetic Modification

One of a pair of HeLa derivatives genetically modified to carry fluorescently tagged histone H2B proteins. H2B is a core component of the nucleosome, where it plays an important role in transcriptional regulation, DNA repair, and other essential cellular functions. One cell line, HeLa H2B-GFP, carries H2B tagged with eGFP. This cell line, HeLa-2FP, carries both a H2B tagged with eGFP and H2B tagged with mCherry. Expression of both proteins enabled study of chromatin compaction in live cells using FLIM-FRET microscopy. The cell lines were published in PNAS in 2019, where they were used to study the remodelling of chromatin architecture at double-strand break sites during DNA repair.

Hazard Group (ACDP)

Hazard Group (ACDP) 2

Applications

References

Original Reference
Jieqiong Lou, Lorenzo Scipioni, Belinda K. Wright, Tara K. Bartolec, Jessie Zhang, V. Pragathi Masamsetti, Katharina Gaus, Enrico Gratton, Anthony J. Cesare, and Elizabeth Hinde
Phasor histone FLIM-FRET microscopy quantifies spatiotemporal rearrangement of chromatin architecture during the DNA damage response
PNAS 2019 Apr 9; 116(15): 7323–7332.
PMID: 30918123

Available Formats

  • Frozen
  • DNA-5µg (100ng/µl)

If use of this culture results in a scientific publication, it should be cited in the publication as: HeLa H2B-2FP (ECACC 21102108).

Unless specified otherwise, at ECACC we routinely handle all of our cell lines at containment level 2 in accordance with the ACDP guidelines (Advisory Committee on Dangerous Pathogens) (UK). All cell cultures have the potential to carry as yet unidentified adventitious agents. It is the responsibility of the end user to ensure that their facilities comply with biosafety regulations for their own country.

 

ACDP Guidance: Biological agents: Managing the risks in laboratories and healthcare premises.

The Culture Collections represent deposits of cultures from world-wide sources. While every effort is made to ensure details distributed by Culture Collections are accurate, Culture Collections cannot be held responsible for any inaccuracies in the data supplied. References where quoted are mainly attributed to the establishment of the cell culture and not for any specific property of the cell line, therefore further references should be obtained regarding cell culture characteristics. Passage numbers where given act only as a guide and Culture Collections does not guarantee the passage number stated will be the passage number received by the customer.

Cultures supplied by Culture Collections are for research purposes only. Enquiries regarding the commercial use of a cell line are referred to the depositor of the cell line. Some cell lines have additional special release conditions such as the requirement for a material transfer agreement to be completed by the potential recipient prior to the supply of the cell line. Please view the Terms & Conditions of Supply for more information.