Bovine Dendritic Cell Growth Kit

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Bovine Dendritic Cell Growth Kit

Product Type
Kits
Product CodeApplicationsDatasheetMSDSPack SizeList PriceQuantity
PBP014KZZ FN 1 ml
Bovine Dendritic cell growth kit (PBP014KZZ) contains a cocktail of biologically active interleukin-4 (IL-4) and granulocyte/macrophage-colony stimulating factor (GM-CSF) that have been premixed at optimal concentrations to induce dendritic cell development from peripheral blood-derived bovine (cattle) monocytes.

Product Details

Target Species
Bovine
Product Form
Mixed recombinant bovine Interleukin-4 and bovine GM-CSF – supplied as a liquid
Preparation
Recombinant cytokines expressed in mammalian Chinese Hamster Ovary (CHO) cells using the pEE14® vector grown in antibiotic free media and USDA-approved dialysed FCS which has been screened for BVDV and virus growth by PCR.
Preservative Stabilisers
None present
Endotoxin Level
<0.5EU/mL

Storage Information

Storage
Store at -20oC only.
Storage in frost-free freezers is not recommended.
This product should be stored undiluted. Avoid repeated freezing and thawing as this may denature this recombinant protein. Should this product contain a precipitate we recommend microcentrifugation before use.
Shelf Life
6 months from date of despatch.

More Information

Acknowledgements
This reagent was produced as part of the BBSRC/SEERAD Immunological Toolbox. The kit development was also supported by the European Community’s Seventh Framework Programme (FP7, 2007-2013), Research Infrastructures action, under the grant agreement No. FP7-228394 (NADIR)
Regulatory
For research purposes only

Applications of Bovine Dendritic Cell Growth Kit

This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visit the antibody protocols page.
Application Name Verified Min Dilution Max Dilution
Functional Assays 1:20
Where this reagent has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the reagent for use in their own system using appropriate negative/positive controls.
Instructions For Use
1. Prepare peripheral blood mononuclear cells (PBMC) from heparinised blood by density gradient centrifugation.

2. Purify CD14+ve cells by labelling PBMC with CD14 mAb and utilise magnetic bead or flow cytometric separation techniques.

3. Resuspend the isolated CD14+ve cells at a concentration of 1x106 cells/ml in tissue culture medium (TCM = RPMI or equivalent + 10% foetal calf serum) containing a final dilution of 1:20 of PBP014KZZ .

4. Add 3ml of cell suspension to each well of a 6 well tissue culture plate.

5. Culture cells in a humidified atmosphere of 5% CO2 in air, at approximately 37oC.

6. Culture cells for 3 days. The cells may then be harvested and used for other procedures including immunophenotyping (as required).

7. If a longer culture period is required the cells must be ‘fed’ with new TCM containing cytokines on day 3:
Carefully remove 1ml spent medium from each well, care is required to avoid disturbing the cells.
Add 1.5ml fresh, pre-warmed TCM containing cytokines at 1:20 to each well and re-culture the DC for required culture period (typically up to 7 days).

8. At the end of the culture period adherent and non-adherent cells can be pooled for use in immunoassays and phenotyped (as required). Adherent cells may require a dissociation step to remove them from the plate.

Useful Reagents Available

Description Product Code Pack Size Applications List Price Quantity
Mouse anti Human CD14:Alexa Fluor® 647 MCA1568A647 100 Tests/1ml F
Mouse anti Human CD14:Alexa Fluor® 700 MCA1568A700 100 Tests/1ml F
Mouse anti Human CD14:Biotin MCA1568B 0.1 mg F
Mouse anti Human CD14:Low Endotoxin MCA1568EL 0.5 mg E F FN WB
Mouse anti Human CD14:FITC MCA1568F 0.1 mg F
Mouse anti Human CD14:RPE-Alexa Fluor® 647 MCA1568P647 100 Tests/1ml F
Mouse anti Human CD14:Pacific Blue® MCA1568PB 100 Tests/1ml F
Mouse anti Human CD14:RPE MCA1568PE 100 Tests F
Mouse anti Bovine CD14:FITC MCA2678F 0.1 mg F
Mouse anti Bovine MHC Class II DQ MCA5655 0.25 mg F IF
Mouse anti Bovine MHC Class II DQ:FITC MCA5655F 0.1 mg F
Mouse anti Bovine MHC Class II DR MCA5656 0.25 mg F
Mouse anti Bovine CD1w2 MCA831G 0.25 mg C F IP

Product Specific References

References for Bovine Dendritic Cell Growth Kit

  1. Hope, J.C et al (2000) Dendritic cells induce CD4+ and CD8+ T-cell responses to Mycobacterium bovis and M. avium antigens in Bacille Calmette Guérin vaccinated and nonvaccinated cattle.
    Scand J Immunol.:52(3):285-91
  2. Myster, F. et al. (2015) Viral semaphorin inhibits dendritic cell phagocytosis and migration but is not essential for gammaherpesvirus-induced lymphoproliferation in malignant catarrhal fever.
    J Virol. 89 (7): 3630-47.
  3. Corripio-Miyar, Y. et al. (2017) 1,25-Dihydroxyvitamin D3 modulates the phenotype and function of Monocyte derived dendritic cells in cattle
    BMC Veterinary Research. 13 (1) [Epub ahead of print].

Further Reading

  1. Werling, D. et al (1999) Involvement of caveolae in the uptake of respiratory syncytial virus antigen by dendritic cells
    Journal of Leukocyte Biology 66: 50-8