Kit Components
1. Anti active Rab5, Mouse Monoclonal Antibody (Catalog No. 26911): One vial – 22 μL (1
    mg/ml) in PBS, pH 7.4, containing 50% glycerol and 0.05% sodium azide. This antibody
    specifically recognizes Rab5-GTP from all vertebrates.
2. Protein A/G Agarose (Catalog No. 30301): One vial – 400 μL of 50% slurry.
3. 5X Assay/Lysis Buffer (Catalog No. 30302): One bottle – 30 mL of 250 mM Tris-HCl, pH 8, 750
    mM NaCl, 50 mM MgCl2, 5 mM EDTA, 5% Triton X-100. 
4. Anti Rab5B, Rabbit Polyclonal Antibody (Catalog No. 21037): One vial – 100 μL (0.2 mg/ml)
    in PBS, pH 7.4, contained 50% glycerol.
5. 100 X GTPγS (Catalog No. 30303): One vial –100 μl at 10 mM, use 5 μL of GTPγS for
    GTP-labeling of 0.5 mL of cell lysate.
6. 100 X GDP (Catalog No. 30304): One vial –100 μl at 100 mM, use 5 μL of GDP for
    GDP-labeling of 0.5 mL of cell lysate. 
Storage
Store all kit components at 4oC until their expiration dates. 
Materials Needed but Not Supplied
1. Stimulated and non-stimulated cell lysates
2. Protease inhibitors
3. 4 °C tube rocker or shaker
4. 0.5 M EDTA, pH8.0
5. 1 M MgCl2
6. 2X reducing SDS-PAGE sample buffer
7. Electrophoresis and immunoblotting systems
8. Immunoblotting wash buffer such as TBST (10 mM Tris-HCl, pH 7.4, 0.15 M NaCl, 0.05%
Tween-20)
9. Immunoblotting blocking buffer (TBST containing 5% Non-fat Dry Milk or 3% BSA)
10. PVDF or nitrocellulose membrane
11. Secondary Antibody
12. ECL Detection Reagents 
Reagent Preparation
? 1X Assay/Lysis Buffer: Mix the 5X Stock briefly and dilute to 1X in deionized water. Just prior to
usage, add protease inhibitors such as 1 mM PMSF, 10 μg/mL leupeptin, and 10 μg/mL aprotinin. 
Note:
For research use only .