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53276S 100 µl (50 tests) $299.00.0
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REACTIVITY SENSITIVITY MW (kDa) Isotype
H Endogenous Mouse IgG3
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Flow Cytometry

Flow cytometric analysis of Ramos cells (blue) and WERI-Rb-1 cells (green) using CD133 (A8N6N) Mouse mAb (Flow Specific) (Alexa Fluor® 647 Conjugate) (solid lines) or a concentration-matched Mouse (MOPC-21) mAb IgG1 Isotype Control (Alexa Fluor® 647 Conjugate) #4843 (dashed lines).

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Flow Cytometry

A. Solutions and Reagents

NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.

  1. 20X Phosphate Buffered Saline (PBS): (#9808) To prepare 1 L 1X PBS: add 50 ml 20X PBS to 950 ml dH2O, mix.
  2. Incubation Buffer: Dissolve 0.5 g Bovine Serum Albumin (BSA) (#9998) in 100 ml 1X PBS. Store at 4°C.

B. Immunostaining

  1. Count cells using a hemacytometer or alternative method.
  2. Aliquot 0.5-1x106 cells into each assay tube (by volume).
  3. Add 2-3 ml Incubation Buffer to each tube and rinse by centrifugation. Repeat.
  4. Resuspend cells in 100 µl of diluted primary antibody (prepared in incubation buffer at the recommended dilution).
  5. Incubate for 1 hr at room temperature.
  6. Wash by centrifugation in 2-3 ml incubation buffer.
  7. Resuspend cells in the appropriate volume of PBS and analyze on flow cytometer.

posted June 2017

Protocol Id: 1504

Product Usage Information

Application Dilutions
Flow Cytometry 1:50

Storage: Supplied in PBS (pH 7.2), less than 0.1% sodium azide and 2 mg/ml BSA. Store at 4°C. Do not aliquot the antibody. Protect from light. Do not freeze.

Specificity / Sensitivity

CD133 (A8N6N) Mouse mAb (Flow Specific) (Alexa Fluor® 647 Conjugate) recognizes endogenous levels of total CD133 protein.


Species Reactivity: Human

Source / Purification

Monoclonal antibody is produced by immunizing animals with cells overexpressing human CD133 protein.

Product Description

This Cell Signaling Technology antibody is conjugated to Alexa Fluor® 647 fluorescent dye and tested in-house for direct flow cytometric analysis in human cells. This antibody is expected to exhibit the same species cross-reactivity as the unconjugated CD133 (A8N6N) Mouse mAb (Flow Specific) #60577.


CD133, also known as Prominin, was first described as a cell surface marker recognized by monoclonal antibody AC133 on putative hematopoietic stem cells (1). Subsequent cDNA cloning indicated that CD133 is a five-transmembrane protein with a predicated molecular weight of 97 kDa. Due to heavy glycosylation, its apparent molecular weight is 130 kDa as determined by SDS-PAGE analysis (2). Besides blood stem cells, CD133 is expressed on and used to isolate other stem cells, including cancer stem cells (3-7). A deletion mutation in CD133 produces aberrant protein localization and may result in retinal degeneration in humans (8).


1.  Yin, A.H. et al. (1997) Blood 90, 5002-12.

2.  Miraglia, S. et al. (1997) Blood 90, 5013-21.

3.  Handgretinger, R. et al. (2003) Ann N Y Acad Sci 996, 141-51.

4.  Monzani, E. et al. (2007) Eur J Cancer 43, 935-46.

5.  O'Brien, C.A. et al. (2007) Nature 445, 106-10.

6.  Ricci-Vitiani, L. et al. (2007) Nature 445, 111-5.

7.  Singh, S.K. et al. (2004) Nature 432, 396-401.

8.  Maw, M.A. et al. (2000) Hum. Mol. Genet. 9, 27-34.


Entrez-Gene Id 8842
Swiss-Prot Acc. O43490


For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
The Alexa Fluor dye conjugates in this product are sold under license from Life Technologies Corporation, for research use only excluding use in combination with DNA microarrays and high content screening (HCS).
This product is provided under an intellectual property license from Life Technologies Corporation. The transfer of this product is contingent on the buyer using the purchased product solely in research, including use with HCS or other automated imaging applications but excluding use in combination with DNA microarrays. The buyer must not sell or otherwise transfer this product or its components for (a) diagnostic, therapeutic or prophylactic purposes; (b) testing, analysis or screening services, or information in return for compensation on a per-test basis; (c) manufacturing or quality assurance or quality control, or (d) resale, whether or not resold for use in research. For information on purchasing a license to this product for purposes other than as described above, contact Life Technologies Corporation, 5791 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@lifetech.com.

53276
CD133 (A8N6N) Mouse mAb (Flow Specific) (Alexa Fluor® 647 Conjugate)