Suitable for use in Flow Cytometry and Western Blot. Other applications not tested.
Recommended Dilution
Flow Cytometry: 10ug/ml. Use 80ul to label 5x10e5 cells. Optimal dilutions to be determined by the researcher.
FACS Protocol
1. Five x10e5 cultured UM-SCC (Squamous Cell Carcinoma) cells were harvested by trypsinization. 2. Five x10e5 cells per tube were washed and pre incubated ~ 5 minutes with 20ul of 250ug/ml of human IgG. 3. Incubate 5 x10e5 cells on ice for 45 minutes with 80ul of C2446-02A at 10ug/ml. 4. Cells were washed twice and incubated with IgG, H&L (FITC) (X-Adsorbed) Pab Gt xMo. 5. Washed 3x, fixed and analyzed on a BD FACS Calibur. 6.. Cells stained positive with a mean shift of 1.78 log10 fluorescent units when compared to a Mouse IgG2a negative control at a similar concentration.
Storage and Stability
May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for at least 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
Immunogen
UM-SCC-1, human squamous cell carcinoma.
Form
Supplied as a liquid in PBS pH 7.5, 0.5mg/ml gentamicin sulfate, <1% Bovine Immunoglobulin.
Purity
Purified by Protein A affinity chromatography.
Specificity
Recognizes the human CD104 molecule at ~ 220kD (non-reduced). Partially blocks binding to laminin.
References
1. K.A. Kimmel and T.E. Carey, Cancer Res (1986) 46: 3614-3623. 2. C. VanWaes, et al, Cancer Res (1991) 51: 2395-2402. 3. Leukocyte Typing V (1995) S.F. Schlossman, et al, (eds.), Oxford University Press, NY. p. 1655-1663, 1667-1668.