Bovine Dendritic Cell Growth Kit
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
- < 0.5 EU/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. - Guarantee
- 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
| 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.
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Useful Reagents Available
| Description | Product Code | Applications | Pack Size | List Price | Quantity |
|---|---|---|---|---|---|
| Mouse anti Human CD14:Alexa Fluor® 647 | MCA1568A647 | F | 100 Tests/1ml |
|
|
| Mouse anti Human CD14:Alexa Fluor® 700 | MCA1568A700 | F | 100 Tests/1ml |
|
|
| Mouse anti Human CD14:Biotin | MCA1568B | F | 0.1 mg |
|
|
| Mouse anti Human CD14:Low Endotoxin | MCA1568EL | E F FN WB | 0.5 mg |
|
|
| Mouse anti Human CD14:FITC | MCA1568F | F | 0.1 mg |
|
|
| Mouse anti Human CD14:RPE-Alexa Fluor® 647 | MCA1568P647 | F | 100 Tests/1ml |
|
|
| Mouse anti Human CD14:Pacific Blue® | MCA1568PB | F | 100 Tests/1ml |
|
|
| Mouse anti Human CD14:RPE | MCA1568PE | F | 100 Tests |
|
|
| Mouse anti Bovine CD14:FITC | MCA2678F | F | 0.1 mg |
|
|
| Mouse anti Bovine MHC Class II DQ | MCA5655 | F IF | 0.25 mg |
|
|
| Mouse anti Bovine MHC Class II DR | MCA5656 | F | 0.25 mg |
|
|
| Mouse anti Bovine CD1w2 | MCA831G | C F IP | 0.25 mg |
|
Product Specific References
References for Bovine Dendritic Cell Growth Kit
-
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 -
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. -
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
-
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
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