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UK Health Security Agency

16-23

16-23

Catalogue No.

99043001

Cell Line Name

16-23

Cell Line Description

Available on request.Please email culturecollections.businessE@phe.gov.uk The cytotoxic monoclonal antibody 16.23 was obtained from a BALB/c mouse immunised with a single i.p. injection of the human melanoma cell line Mel Juso. Mouse spleen cells were removed 3 days after immunisation and fused with the myeloma P3x63Ag8.653. The Juso B cell line was typed as HLA A1,2; B7,8; CW- ; DR2,3. The monoclonal antibody 16.23 detects the conformation-dependant DR3 epitope but was also reported to react with some HLA-Dw/DRw6 individuals. Together with antibodies S2 (ECACC catalogue no. 99043002) and S5 (ECACC catalogue no. 99061827) a useful tool for the biochemical dissection of DRw6 related epitopes has been establised. It has been analysed in the 9th International Histocompatibility Workshop (9W929). The hybridoma line has been eradicated from mycoplasma using Cibrobay before depositing at ECACC.

Characteristics

Myeloma

P3X63 Ag8.653

Immunogen

Human melanoma cell line Mel Juso

Immunological Donor

BALB/c mouse spleen

Antibody Isotype

IgG3

Morphology

Spherical

Tissue of Origin

Hybrid

Culture Conditions

Cell Type

Hybridoma

Subculture Routine

Maintain cultures between 3-9 x 100,000 cells/ml; 5% 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 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 + 2mM glutamine + 1mM NaP + 10% FBS

Growth Mode

Suspension

Additional Info

Depositor

Dr. JP Johnson, Institute for Immunology, University of Munich

Country of Origin

Germany

Applications

Immunohistochemistry, Cytotoxicity, Immunoprecipitation

Assays

Immunogluorescence, Immunoperoxidase on melanoma cell lines

Hazard Group (ACDP)

2

References

References

J. Exp Med. 1982;156:104,Immunity 1996; 4:87, J Immunol 1995; 154:2545,J Immunol 1992; 149: 754, J Immunol 148:2586, Immun Today 1989; 10:305, Hum Immunol 1986; 16:69, Eur J Immunol 1985; 15:1239, J Immunol 1985; 135:2393, J Exp Med 1985; 162:1193,

Available Formats

  • Frozen

If use of this culture results in a scientific publication, it should be cited in the publication as: 16-23 (ECACC 99043001)

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.