Stemgent® Affinity Purified anti-Human SSEA-4 Antibody详细信息：
Product Overview
The MC-813-70 monoclonal antibody reacts with stage-specific embryonic antigen-4 (SSEA-4). SSEA-4 is expressed on teratocarcinoma (embryonic carcinoma, EC), human embryonic germ (EG) cells, and human embryonic stem (ES) cells, as well as on a subset of mesenchymal stem cells, erythrocytes, and the erythromyeloid leukemia cell line K562. SSEA-4 is widely used to characterize human ES cells and to monitor their differentiation, as the expression of SSEA-4 on human ES cells is downregulated upon differentiation. In the mouse, SSEA-4 is expressed on oocytes and early cleavage-stage embryos, but not on EC or ES cells.
Product Specifications
Stemgent® Affinity Purified anti-Human SSEA-4 AntibodyImunogen Human embryonic carcinoma (EC) cell line 2102 Ep
Reactivity Human
Clone MC-813-70
Isotype Mouse IgG3, κ
Size 50 μg (0.1 ml at 0.5 mg/ml)
Formulation Phosphate-buffered solution, pH 7.2, containing ≤0.09% sodium azide.
Preparation The antibody was purified by affinity chromatography.
Storage Store undiluted at 4°C. Do not freeze.
Stability Stable for 6 months from date of receipt when stored as directed.
Quality Control Tested by flow cytometry and immunocytochemistry to ensure product quality.
Recommended Usage 0.25 μg per 1x106 viable cells for flow cytometry and 5 μg/ml for immunocytochemistry.
General Protocol for Immunofluorescent Staining Using Flow Cytometry
Isolate and dissociate cells to a single cell suspension. Collect cells in a 15 ml conical tube.
Centrifuge the cell suspension at 300 x g for 5 minutes at 4°C. Resuspend the cell pellet to a concentration of 0.2x107 to 1x107 cells per ml in PBS. Keep cells on ice.
For each sample, add 100 μl of the cell suspension to a 4 ml roundbottom tube and add the appropriate amount (10-20 μl) of the purified antibody.
Stemgent® Affinity Purified anti-Human SSEA-4 AntibodyIncubate at 4°C for 30 minutes to 1 hour.
Wash the cells by adding PBS to bring cell suspension to 4 ml. Centrifuge at 300 x g for 5 minutes at 4°C.
Aspirate and agitate the cell pellet. Then add the appropriate amount of FITC-conjugated anti-mouse IgM antibody to each sample.
Incubate at 4°C for 30 minutes to 1 hour.
Add PBS to bring cell suspension to 4 ml. Centrifuge at 300 x g for 5 minutes at 4°C.
Aspirate and agitate the cell pellet. Add 10 &muL (0.25 &mug) of 7- Aminoactinomycin D (cell viability dye) to each sample and incubate for 20 minutes at 4°C.
Add appropriate volume of PBS to each tube. The cells are ready to be analyzed by flow cytometry.
General Protocol for Immunocytochemistry
Seed appropriate number of ES cells in a 12-well plate on mouse embryonic fibroblast (MEF) feeder cells. Culture cells in a 37°C incubator with 5% CO2 for 3-4 days.
Wash the cells gently with PBS 3 times.
Fix the cells with 500 μl fixative solution for 20 minutes at room temperature.
Wash the cells with PBS 3 times.
Block non-specific binding with 500 μl appropriate blocking buffer for 1 hour at room temperature.
Incubate the cells with 200 μl of the antibody overnight at 4°C.
Wash the cells with PBS 3 times.
Incubate with 200 μl of appropriate conjugated secondary antibody for 2 hours at room temperature. Avoid exposing the plate to light.
Wash cells with PBS 3 times.
During the last wash, add DAPI stain to a final concentration at 1 μg/ml for 5 minutes.
After the final wash with PBS, the cells are ready to be analyzed using a fluorescent microscope.
Notice to Purchaser Purchaser represents and warrants that it will use Phycoerythrin (PE) antihuman SSEA-4 purely for research purposes, not for diagnostic use, not for resale, and not for use in humans or veterinary applications. Stemgent will not be held responsible for patent infringement or other violations that may occur with the use of our products.
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