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To Purchase # 60577S

60577S 100 µl (50 tests) $239.00
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REACTIVITY SENSITIVITY MW (kDa) Isotype
H Endogenous Mouse IgG3
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Flow Cytometry

Flow cytometric analysis of live Ramos cells (blue) and Caco-2 cells (green) using CD133 (A8N6N) Mouse mAb (Flow Specific).

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Flow Cytometry

A. Solutions and Reagents

B. Immunostaining

NOTE: Account for isotype matched controls for monoclonal antibodies or species matched IgG for polyclonal antibodies. Count cells using a hemacytometer or alternative method.

  1. Collect cells by centrifugation and aspirate supernatant.
  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 Incubation Buffer per assay tube.
  5. Block in Incubation Buffer for 10 minutes on ice.
  6. Add the unconjugated primary antibody at the appropriate dilution to the assay tubes (see individual antibody data sheet for the dilution recommendations).
  7. Incubate for 1 hour on ice.
  8. Rinse as before in Incubation Buffer by centrifugation.
  9. Resuspend cells in fluorochrome-conjugated secondary antibody* diluted in total volume Incubation Buffer at the recommended dilution.
  10. Incubate for 30 minutes on ice.
  11. Rinse as before in Incubation Buffer by centrifugation.
  12. Resuspend cells in 0.5 ml PBS and analyze on flow cytometer.

posted Februay 2011

Protocol Id: 58

Product Usage Information

Application Dilutions
Flow Cytometry 1:50

Storage: Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Specificity / Sensitivity

CD133 (A8N6N) Mouse mAb (Flow Specific) 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.

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.

60577
CD133 (A8N6N) Mouse mAb (Flow Specific)