CTLA4 Mouse Monoclonal Antibody [Clone ID: A3.4H2.H12]

CAT#: AM31866PU-N

CTLA4 mouse monoclonal antibody, clone A3.4H2.H12, Purified

Specifications

Product Data

• Clone Name: A3.4H2.H12
• Applications: FC
• Recommended Dilution: Flow Cytometry.
• Reactivities: Human
• Host: Mouse
• Isotype: IgG2a
• Clonality: Monoclonal
• Immunogen: Activated T cells from Balb/c mouse.
  Fusion Partner: SP 2/0 myeloma.
• Specificity: Recognizes Human CTLA-4 (CD152).
  Other species not tested.
• Formulation: PBS containing 0.02% Sodium Azide as preservative
  State: Purified
  State: Liquid purified IgG fraction
• Concentration: lot specific
• Purification: Affinity Chromatography on Protein G
• Conjugation: Unconjugated
• Storage: Store undiluted at 2-8°C for one month or (in aliquots) at -20°C for longer.
  Avoid repeated freezing and thawing.
• Stability: Shelf life: one year from despatch.
• Gene Name: cytotoxic T-lymphocyte associated protein 4
• Database Link:
  Entrez Gene 1493 Human
  UniProt ID P16410
• Background: Human CTLA-4 (also known as Cytotoxic T lymphocyte-associated antigen-4, CD152) is a structural homologue of T cell co-stimulatory receptor CD28. CTLA-4 binds to the same ligands (CD80 and CD86) as CD28 but with much higher avidity. It is expressed at very low levels on the cell surface of activated T cells, and is found primarily in the post-Golgi or endosomal compartments of the cell.
  There is a high degree of overall homology between human and mouse CTLA-4 (>70%), and the cytoplasmic domains of the human, mouse and rabbit sequences are completely conserved. CD152 antibodies have shown both stimulatory and negative regulatory roles in functional experiments.
• Synonyms: CTLA-4
• Note: Protocol: FLOW CYTOMETRY ANALYSIS:
  1. Prepare cell suspension in Media A. For cell preparations, deplete the red blood cell population with Lympholyte®-H cell separation medium.
  2. Wash 2 times.
  3. Resuspend cells to 1x10e6 cells in approximately 50 μl Media A in a microcentrifuge tube (i.e. 50 μl of cells resuspended to 2x10e7 cells/ml).
  (the contents of 1 tube represent 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-mouse IgG (H+L)) at 1/500 dilution.
  9. Incubate 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 phosphate buffered saline. (This stains dead cells by intercalating 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 mls).

Reference Data

• Protein Families: Druggable Genome, Transmembrane
• Protein Pathways: Autoimmune thyroid disease, Cell adhesion molecules (CAMs), T cell receptor signaling pathway