Cd80 Rat Monoclonal Antibody [Clone ID: RMMP-1]

CAT#: AM31878PU-N

Cd80 rat monoclonal antibody, clone RMMP-1, Purified

Specifications

Product Data

• Clone Name: RMMP-1
• Applications: FC
• Recommended Dilution: Flow Cytometry (See Protocols).
• Reactivities: Mouse
• Host: Rat
• Isotype: IgG2a
• Clonality: Monoclonal
• Specificity: This antibody recognises the murine CD80 cell surface protein, also known as B7-1.
• Formulation: PBS containing 0.09% Sodium Azide as preservative and EIA grade BSA to bring total protein concentration to 4-5 mg/ml
  State: Purified
  State: Liquid purified IgG fraction
• Concentration: 0.2 mg/ml
• Gene Name: Mus musculus CD80 antigen (Cd80)
• Database Link:
  Entrez Gene 12519 Mouse
• Background: CD80 is a member of the Ig superfamily, along with CD86 (B7-2), participates in T cell costimulation via interactions with CD28 and CTLA-4. CD80 is constitutively expressed on dendritic cells, monocytes, and peritoneal macrophages; and it is inducible on B cells by various means, including activation by LPS, IL-4, and the cross-linking of surface Ig. Expression of CD80 is greatly enhanced on splenic B cells following activation by LPS, with peak expression occurring between 48 and 72 hours. It has been reported that activation of purified B cells with LPS can induce CD80 expression in as few as 18 hours.
• Synonyms: CD28LG, CD28LG1, LAB7, BB1, B7.1, B7-1
• Note: Protocol: FLOW CYTOMETRY ANALYSIS:
  NOTE: Each lot is tested using freshly harvested mouse splenocytes and mouse splenocytes cultured for 72 hrs with LPS (30 ug/ml).
  
  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 ~0.25 μg* of AM31878PU-N.
  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 FITC Goat anti-Rat IgG (H+L) secondary antibody 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 mls).