COMBI IC Reagent: IgG Negative Control (FITC) and IgG Negative Control (PE)


Catalogue number: GIC-201

CloneVI-AP and VI-AP
IsotypeIgG1
Product Type Negative Control
Units1 ml
HostMouse
Application Flow cytometry
Background
This ready to use Negative Control reagent contains a combination of fluorescein and phycoerythrin conjugated mouse immunoglobulin molecules of IgG1 isotype, which have been selected on the basis of their binding characteristics: no specific binding to human intracellular or cell surface antigens, same low range of nonspecific binding to human leukocytes as other COMBI-IC-Reagents. Like all other COMBI-IC-reagents, this reagent should be used in combination with our FIX&PERM Cell Permeabilization Kit (Cat.No. GAS-002) These isotype control IgG1 are suitable as negative controls to be used in combination with COMBI-IC reagents for the: - Enumeration of Myeloid Cells - Analysis of Myeloid Differentiation Stage - Enumeration of B-cells and Precursors - Enumeration of T-cells and Precursors - Analysis of Leukemia Cells - Analysis of Immunodeficiency States Results must be put within the context of other diagnostic tests as well as the clinical history of the patient by a certified professional before final interpretation.

Product
1ml of FITC- and PE-conjugated VI-AP in PBS pH 7.2, 1% BSA, and 0.05% NaN3, approximately 50 tests.

Applications
Permeabilization and Staining Procedure - In combination with our Permeabilization Kit FIX&PERM (Cat. No. GAS-002) intracellular isotype controls can be easily stained in cell suspensions. - For each sample to be analyzed add 50 µl of whole blood, bone marrow or mononuclear cell suspension in a 5ml tube - Add 100 µl of Reagent A (Fixation Medium, stored and used at room temperature) - Incubate for 15 minutes at room temperature - Add 5 ml phosphate buffered saline and centrifuge cells for 5 minutes at 300 g - Remove supernatant and add to cell pellet 100 µl Reagent B (Permeabilization Medium) and 20 µl of COMBI-IC Negative Control antibody conjugate. - Vortex at low speed for 1-2 seconds - Incubate for 15 minutes at room temperature - Wash cells with phosphate buffered saline as described above - Remove supernatant and resuspend cells in sheath fluid for immediate analysis or resuspend cells in 0.5 ml 1.0 % formaldehyde and store them at 2-8°C in the dark. Analyze fixed cells within 24 hours.

Specificity
VI-AP reacts with calf intestine alkaline phosphatase and does not show cross-reactivity with human proteins.

Storage
Nordic-MUbio monoclonal antibody reagents contain optimal concentrations of affinity-purified antibody. For stability reasons this monoclonal antibody solution contains sodium azide. These reagents should be stored at 2-8°C (DO NOT FREEZE!) and protected from prolonged exposure to light. Stability of the reagent: Please refer to the expiry date printed onto the vial. The use of the reagent after the expiration date is not recommended.

References
1. J. L. Clarke, W. Watkins, J Biol Chem 271, 10317 (1996).
2. R. N. Knibbs et al., J Cell Biol 133, 911 (1996).
3. B. Kniep et al., J Biochem (Tokyo) 119, 456 (1996).
4. A. J. Wagers, L. M. Stoolman, R. Kannagi, R. Craig, G. S. Kansas, J Immunol 159, 1917 (1997).
5. M. Noguchi, N. Sato, H. Sugimori, K. Mori, K. Oshimi, Leuk Res 25, 847 (2001).
6. W. M. Watkins, J. L. Clarke, Adv Exp Med Biol 491, 231 (2001).

Caution
For professional users only. This reagent contains sodium azide. To avoid the development of hazardous conditions, reagents containing azide should be diluted in running water prior to be discarded. Similar to the work with other biological products, proper handling procedures are recommended.