Cell Culture

1. Introduction to cell culture

2. Media preparation

3. DMEM

4. Dialysis of FBS

5. Splitting

6. Hemacytometer

7. Coulter counter

8. Trypan blue staining

9. Freeze/thaw

10. Freezing cell lines

11. Thawing cells

12. HeLa S3 cell culture

13. Xenopus oocyte microinjection

14. Preparation of extracts from human placenta

15. Chicken blood injections

16. Sacrifice of adult hen

17. Bleeding eggs

18. Chicken embryonic fibroblasts

Cell Transfection

1. CaPO4 method

2. 3% CO2 method

3. Ecogpt

4. Trypsinization transfection

5. Polyfect, Lipofectamine, and Effectene

6. Cell sorting for transfected cells: MACs protocol

Establishing Stable Cell Lines

1. Calcium Chloride Transfection

2. Lipofectamine Transfection (Gen’s Protocol)

Radiolabeling

1. 32P orthophosphate and 35S methionine/cysteine

2. Metabolic labeling

Cell Growth/Death Assays

1. Cell Death ELISA

2. MTT

3. TUNEL

4. Drug Assays

Luciferase Assays

1. Luminometer operation

2. Single luciferase

3. Dual luciferase

4. 96-well plate format

RNAi – mammalian cells

Cell Imaging

1. Intro to immunofluorescence

2. Method

3. FRET

Transferrin Uptake using Fluorescence Microscopy