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UK Health Security Agency

JB1

JB1

Catalogue No.

01091808

Cell Line Name

JB1

Cell Line Description

To produce this hybridoma mice transgenic HLA-B27 and human beta-2 microglobulin were immunised with a preparation of HFE made in recombinant bacteria, refolded with human beta-2 microglobulin. Characterisation beyond data in B.J. Haematol 103, 931, 1998 shows that JB1 reacts predominantly with aggregated denatured HFE produced in bacteria, but does not bind folded recombinant HFE produced in CHO cells. In comparison with polyclonal sera raised to the C terminus or the alpha 3 region of HFE JB1 stains frozen sections of the tissues well, but reacts very weakly with cells transfected with the HFE gene in vitro (usually only visible by confocal microscopy). JB1 does not stain paraffin fixed sections and will not work in western blots.

 

Characteristics

Myeloma

PS-NS-1 Ag4.1

Immunogen

Bacterial recombinant HFE protein

Immunological Donor

(BALB/c x B27) female transgenic mouse

Antibody Isotype

IgG3

Morphology

Spherical

Tissue of Origin

Hybrid

Culture Conditions

Cell Type

Hybridoma

Subculture Routine

Maintain cells at 3-5 x 100,000 cells/ml which is usually a split ratio of 1:2. Saturation density is 5 x 100,000 to 1 x 1000,000. 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 5-10ml of prewarmed 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 10 µ cells/ml. Place culture flask flat and observe regularly until viable proliferating cells are seen. Often hybridoma cultures may benefit from being resuspended 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

RPMI 1640, 10% FBS, 1% HAT

Growth Mode

Suspension

Additional Info

Depositor

Dr Judy Bastin, Weatherall Institue of Molecular Medicine, John Radcliffe Hospital, Oxford. OX3 9DS

Country of Origin

United Kingdom

Assays

Elisa, immunohistrochemistry

Hazard Group (ACDP)

2

References

References

British Journal of Haematology 1998, 103, 931-94

Available Formats

  • Frozen

If use of this culture results in a scientific publication, it should be cited in the publication as: JB1 (ECACC 01091808)

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.