Cr1l Mouse Monoclonal Antibody [Clone ID: 512]

CAT#: AM31835PU-N

Cr1l mouse monoclonal antibody, clone 512, Purified

Product Images

Cell Source: Bone Marrow Percentage of cells stained above control: 98.7%

Specifications

Product Data

• Clone Name: 512
• Applications: FC, IHC
• Recommended Dilution: Flow Cytometry.
  Immunohistochemistry on aceton-fixed frozen sections.
• Reactivities: Rat
• Host: Mouse
• Isotype: IgG1
• Clonality: Monoclonal
• Immunogen: Erythrocytes from a C3 mutated rat
• Specificity: Purified Anti-Rat Crry monoclonal antibody (Clone: 5I2) is a rat-specific membrane complement regulator that can inhibit both antibody-induced classical pathway and alternative pathway complement activation.
  Crry plays a more dominant role than DAF in regulating the alternative pathway of complement and in preventing spontaneous complement damage of rat erythrocytes, whereas DAF and Crry are both expressed and are equally effective in preventing antibody-induced runaway complement activation on rat erythrocytes.
• 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: Rattus norvegicus complement C3b/C4b receptor 1 like (Cr1l), transcript variant 3
• Database Link:
  Entrez Gene 54243 Rat
• Synonyms: Antigen 5I2
• Note: Protocol: FLOW CYTOMETRY ANALYSIS:
  1. Prepare cell suspension in Media A. For cell preparations, deplete the red blood cell population with Lympholyte®-R 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 0.1 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).
  
  Results:
  Tissue Distribution by Flow Cytometry Analysis:
  Rat Strain: Wister
  Cell Concentration: 1x10e6 cells per test
  Antibody Concentration Used: 0.1μg/10e6 cells
  Isotypic Control: Purified Mouse IgG1
  Cell Source:
  Whole Blood 44.6%
  Bone Marrow 98.7%
  Spleen 77.8%