This antibody reacts with Human CD123 antigen (IL-3 receptor alpha-chain).
Cross-Reactivity (Details)
Species reactivity (tested):Human.
Purification
Protein-A Sepharose Chromatography.
Immunogen
COS cells overexpressing IL-3 receptor alpha-chain. Remarks: Hybridoma was established by fusion of mouse myeloma cell NS-1 with Balb/c mouse splenocyte
IL3RA
Reactivity: Human
FACS, ELISA
Host: Rabbit
Monoclonal
DM31
unconjugated
Application Notes
Flow Cytometry: 20 μL (Ready for Use). Positive Control: Transfectant cells. Detailed procedure is provided in the following Protocols. Other applications not tested. Optimal dilutions are dependent on conditions and should be determined by the user.
Protocol
Flow Cytometric Aanalysis for Adherent CellsProtocol 1We usually use Fisher tubes or equivalents as reaction tubes for all steps after 2). 1) Detach the cells from culture dish by trypsinization. *Excessive trypsinization mayreduce the antigenicity. 2) Wash the cells 3 times with washing buffer [PBS containing 2% fetal calf serum (FCS)and 0. 1% NaN3]. 3) Resuspend the cells with washing buffer (5x10e6 cells/mL). 4) Add 50 µL of the cell suspension into each tube, and centrifuge at 500 x g for 1 minute atroom temperature (20~25°C). Remove supernatant by careful aspiration. 5) Add 10 µL of normal goat serum containing 1 mg/mL normal human IgG and 0. 1% NaN3to the cell pellet after tapping. Mix well and incubate for 5 minutes at room temperature(20~25°C)6) Add 20 µL of the PE labeled CD123 monoclonal antibody (6H6). Mix well, and incubatefor 30 minutes at room temperature. 7) Add 1 mL of the washing buffer followed by centrifugation at 500 x g for 1 minute at roomtemperature (20~25°C). Remove supernatant by careful aspiration. 8) Resuspend the cells with 500 µL of the washing buffer and analyze by a flow cytometer. Positive Control for Flow Cytometry: transfectant
Restrictions
For Research Use only
Format
Liquid
Buffer
PBS containing 1 % BSA as stabilizer and 0.09 % Sodium Azide as preservative.
Preservative
Sodium azide
Precaution of Use
This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
CD123 is the IL-3-specific component of the IL-3 receptor. CD123 binds IL-3 only in combination with CDw131, the Synonyms: IL-3R-alpha, IL3 receptor alpha, Interleukin-3 receptor subunit alpha