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UK Health Security Agency

4.6.5a114K

4.6.5a114K

Catalogue No.

93062218

Cell Line Name

4.6.5a114K

Cell Line Description

The antibody is directed against a 144kDa protein present predominantly on the apical, but also on the basolateral surfaces of Madin-Darby canine kidney cells of the MDCK cell line.

Characteristics

Myeloma

Sp2/0 Ag14

Immunogen

Basolateral membrane fraction from dog intestine

Immunological Donor

BALB/c mouse spleen

Antibody Isotype

IgG1κ

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

EMEM (EBSS) + 2mM glutamine + 1% Non Essential Amino Acids (NEAA) + 10% Foetal Bovine Serum (FBS)

Growth Mode

Suspension

Additional Info

Depositor

Dr K Simons, European Molecular Biology Laboratory, Heidelberg

Country of Origin

Germany

Applications

Study of cell surface polarity development

Assays

Immunofluorescence, immunoprecipitation

Hazard Group (ACDP)

2

References

References

EMBO J 1984;11:2687; J Cell Biol 1986;102:457

Available Formats

  • Frozen

If use of this culture results in a scientific publication, it should be cited in the publication as: 4.6.5a114K (ECACC 93062218)

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.