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Ordering Information
- : ES-011-B
- : 500 mL
EmbryoMax® ES Cell Qualified Fetal Bovine Serum, New Zealand origin
Description:
EmbryoMax® ES Cell Qualified Fetal Bovine Serum, New Zealand origin
Qty/Pk:
500 mL
Product Overview:
The maintanence, expansion, and assay of many stem cell and precursor cell cultures relies on media rich in a variety of factors, often supplied by fetal bovine serum (FBS). Such sera must be carefully controlled and tested. Millipore utilizes sera that are approved by the United States Department of Agriculture (USDA) to ensure that sera are free of diseases such as Bovine Spongiform Encephalopathy (BSE). A Certificate of Analysis is available for each lot of EmbryoMax ES Cell Qualified FBS, identifying the USDA-approved country of origin.
Key Applications:
Stem Cell Culture
Usage Statement:
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Species:
Fetal Bovine
Quality Assurance:
Each lot is tested and qualified for use with Embryonic Stem Cells.
Sera tested: Positive Control FBS; Negative Control FBS; FBS Lots to be tested.
Cells tested: ES Cells derived from strain 129/SVEV
Experiment:
Day 1:
1. Treat feeder cells with 10μg/ml of Mitomycin C for 2.5 hours at 37°C.
2. Remove Mitomycin C medium and wash cells 3X with FBS.
3. Add Trypsin / EDTA and incubate at 37°C for 5 minutes.
4. Add 2 volumes of medium with serum, pipet cells, remove sample to count.
5. Pellet cells at 1000 rpm, 5 minutes, 4°C.
6. Resuspend cells at 2.25x105 cells/ml in medium without serum. 7. Add 1.8 ml of feeder cell suspension/well of 6 well dish treated with 0.1% gelatin.
8. Add 0.2 ml of test serum to each 10% test well and 0.6 mls of test serum to each 30% test well. Incubate at 37°C, 5%CO2.
9. Remove medium from ES cells (growing on PMEF feeder cells).
10. Wash with PBS. Add Trypsin/ EDTA and incubate at 37°C for 5 minutes.
11. Add 2 volumes of medium with serum, pipet cells, remove sample to count.
12. Pellet cells at 1000 rpm, 5 minutes, 4°C.
13. Resuspend cells at 2 x 104 cells/ml in medium without serum.
14. Add 25μL of ES cell suspension to each well (~500 cells / well) and incubate at 37°C, 5%CO2.
Day 5
1. Examine plates under microscope.
2. Feed one half of experiment fresh medium and serum.
Day 9
1. Examine plates by eye and under microscope.
2. Record Observations and conclusions.
3. Remove media, wash cells with PBS, fix with 100% methanol, stain with Giemsa stain, rinse with water, and air dry. Examine plates by eye and under microscope.
Sera tested: Positive Control FBS; Negative Control FBS; FBS Lots to be tested.
Cells tested: ES Cells derived from strain 129/SVEV
Experiment:
Day 1:
1. Treat feeder cells with 10μg/ml of Mitomycin C for 2.5 hours at 37°C.
2. Remove Mitomycin C medium and wash cells 3X with FBS.
3. Add Trypsin / EDTA and incubate at 37°C for 5 minutes.
4. Add 2 volumes of medium with serum, pipet cells, remove sample to count.
5. Pellet cells at 1000 rpm, 5 minutes, 4°C.
6. Resuspend cells at 2.25x105 cells/ml in medium without serum. 7. Add 1.8 ml of feeder cell suspension/well of 6 well dish treated with 0.1% gelatin.
8. Add 0.2 ml of test serum to each 10% test well and 0.6 mls of test serum to each 30% test well. Incubate at 37°C, 5%CO2.
9. Remove medium from ES cells (growing on PMEF feeder cells).
10. Wash with PBS. Add Trypsin/ EDTA and incubate at 37°C for 5 minutes.
11. Add 2 volumes of medium with serum, pipet cells, remove sample to count.
12. Pellet cells at 1000 rpm, 5 minutes, 4°C.
13. Resuspend cells at 2 x 104 cells/ml in medium without serum.
14. Add 25μL of ES cell suspension to each well (~500 cells / well) and incubate at 37°C, 5%CO2.
Day 5
1. Examine plates under microscope.
2. Feed one half of experiment fresh medium and serum.
Day 9
1. Examine plates by eye and under microscope.
2. Record Observations and conclusions.
3. Remove media, wash cells with PBS, fix with 100% methanol, stain with Giemsa stain, rinse with water, and air dry. Examine plates by eye and under microscope.
Stem Cell Type:
- Mouse Embryonic Stem Cells
- Induced Pluripotent Stem Cells
Stem Cell Workflow Stage:
Maintenance & Expansion
Brand Family:
Specialty Media
Presentation:
Liquid
Storage Conditions:
Store at -20°C up to 2 years from date of receipt.
Cell Culture Reagent Type:
- Stem Cell Reagents
- Cell Culture Supplements
Product Name:
EmbryoMax® ES Cell Qualified Fetal Bovine Serum, New Zealand origin