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UK Health Security Agency

L3B2

L3B2

Catalogue No.

19072505

Cell Line Name

L3B2

Cell Line Description

L3B2 was developed by immunising mice with Trypanosoma brucei CaCl2 precipitated flagella preparation.

Mouse spleen cells were fused with the myeloma X63-Ag8.653.

Antibody: Tested at ECACC and found to be IgG1

Characteristics

Myeloma

X63-Ag8.653

Immunogen

L3B2 was developed by immunising mice with Trypanosoma brucei CaCl2 precipitated flagella prep

Antibody Isotype

IgG1

Morphology

Spherical

Tissue of Origin

Hybrid

Culture Conditions

Cell Type

Hybridoma

Subculture Routine

Maintain cultures between 1-9 x 10⁵ cells/ml; 5 to 8% CO2; 37°C.

When recovering hybridoma cultures from frozen it is not unusual for growth to be slower than expected initially and there may be an observed decrease in viability. Establishment of an actively proliferating culture may take up to 2 weeks.
On resuscitation a centrifugation step to remove the cryoprotectant is essential. Rapidly thaw the frozen ampoule in a water bath at 37°C for 1-2 minutes. Transfer the contents to a centrifuge tube and slowly add pre-warmed growth media+. Remove a sample for counting. Centrifuge at 100 x g for 2-3 minutes to pellet cells and seed at a relatively high density of 5-7 x 10⁵ cells/ml.

Place culture flask flat and observe regularly until viable proliferating cells are seen.

Often hybridoma cultures may benefit from being re-suspended with media supplemented with 20% FBS in the early critical stage of culture establishment immediately post resuscitation. Once the culture is established in culture the FBS can be reduced to 10%.

Culture Medium

DMEM + 4mM glutamine + 10% Foetal Bovine Serum (FBS)

Growth Mode

Suspension

Additional Info

Depositor

Keith Gull, Sir William Dunn School of Pathology

Country of Origin

United Kingdom

Applications

Immunoelectron microscopy, immunofluorescence and Western Blot.

Hazard Group (ACDP)

1

References

References

Kohl, L et al. “Assembly of the paraflagellar rod and the flagellum attachment zone complex during the Trypanosoma brucei cell cycle.” The Journal of eukaryotic microbiology vol. 46,2 (1999): 105-9. doi:10.1111/j.1550-7408.1999.tb04592.x PMID: 10361731

 

Gadelha, Catarina et al. “Relationships between the major kinetoplastid paraflagellar rod proteins: a consolidating nomenclature.” Molecular and biochemical parasitology vol. 136,1 (2004): 113-5. doi:10.1016/j.molbiopara.2004.03.006 PMID: 15138073

Available Formats

  • Frozen

If use of this culture results in a scientific publication, it should be cited in the publication as: L3B2 (ECACC 19072505)

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.