Ly6a Rat Monoclonal Antibody [Clone ID: CT-6A/6E]

CAT#: AM31883PU-N

Ly6a rat monoclonal antibody, clone CT-6A/6E, Purified

Specifications

Product Data

• Clone Name: CT-6A/6E
• Applications: FC
• Reactivities: Mouse
• Host: Rat
• Isotype: IgG2b
• Clonality: Monoclonal
• Specificity: This monoclonal antibody recognizes Sca-1 (Ly-6A.2/6E.1), a cell surface antigen used in the identification of hematopoietic stem cells.
• Formulation: PBS containing 0.02% sodium azide (NaN3) as preservative
  State: Purified
  State: Liquid purified Ig fraction
• Concentration: 1,0 mg/ml
• Purification: Affinity chromatography on Protein G
• Gene Name: Mus musculus lymphocyte antigen 6 complex, locus A (Ly6a), transcript variant 3
• Database Link:
  Entrez Gene 110454 Mouse
• Background: Ly6A/E is a member of the Ly-6 antigen family. The Thy-1lo, Lin- (lineage-negative, not expressing B220, Gr-1, Mac-1, CD4 or CD8), Sca-1+ population of bone marrow cells are highly purified, perhaps homogenous, pluripotent stem cells. This antigen is also present on various other tissues. Specific staining of the parenchymal cells can be demonstrated in thymus, spleen and kidney where as only vasculature reacts with anti-Sca-1 in brain, heart and liver (and possibly in lung). Also, Sca-1 is a T cell activation antigen, as surface expression of the antigen increases upon Con A activation of T lymphocytes. Sca-1 appears to have a molecular mass of 8 kDa under non-reducing conditions and 18 kDa under reducing conditions, indicating the presence of intra-chain disulfide bonds.
• Synonyms: Lymphocyte antigen 6A-2/6E-1, Ly-6A.2/Ly-6E.1, T-cell-activating protein, TAP, Stem cell antigen 1, SCA-1
• Note: Protocol: FLOW CYTOMETRY ANALYSIS:
  Method:
  1. Prepare a cell suspension in media A. For cell preparations, deplete the red blood cell population with Lympholyte®-M cell separation medium.
  2. Wash 2 times.
  3. Resuspend the cells to a concentration of 2x10e7 cells/ml in media A. Add 50 μl of this suspension to each tube (each tube will then contain 1x10e6 cells, representing 1 test).
  4. To each tube, add 1.0 μg of this antibody per 10e6 cells.
  5. Vortex the tubes to ensure thorough mixing of antibody and cells.
  6. Incubate the tubes for 30 minutes at 4°C.
  7. Wash 2 times at 4°C.
  8. Add 100 μl of secondary antibody (FITC Goat anti-rat IgG (H+L)) at 1/500 dilution.
  9. Incubate the tubes at 4°C for 30-60 minutes. (It is recommended that the tubes are protected from light since most fluorochromes are light sensitive).
  10. Wash 2 times at 4°C in media B.
  11. Resuspend the cell pellet in 50 μl ice cold media B.
  12. Transfer to suitable tubes for flow cytometric analysis containing 15 μl of propidium iodide at 0.5 mg/ml in PBS. This stains dead cells by intercalating in DNA.
  Media:
  A. Phosphate buffered saline (pH 7.2) + 5% normal serum of host species + sodium azide (100 μl of 2M sodium azide in 100 mls).
  B. Phosphate buffered saline (pH 7.2) + 0.5% Bovine serum albumin + sodium azide (100 μl of 2M sodium azide in 100 ml
  Results:
  Tissue Distribution by Flow Cytometry Analysis:
  Mouse Strain: Balb/C
  Cell Concentration: 1 x 10e6 cells per test
  Antibody Concentration Used: 1.0 μg/10e6 cells
  Isotypic Control: Purified Rat IgG2b