Cr1l Mouse Monoclonal Antibody [Clone ID: 512]

CAT#: AM31835BT-N

Cr1l mouse monoclonal antibody, clone 512, Biotin

Product Images

Cell Source: Spleen Percentage of cells stained above control: 96.7%

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: 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) and EIA grade BSA as a stabilizing protein to bring total protein concentration to 4-5 mg/ml.
  Label: Biotin
  State: Liquid purified Ig fraction
• Concentration: 0,1 mg/ml
• Conjugation: Biotin
• 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:
  Method:
  1. Prepare a cell suspension in media A. For cell preparations, deplete the red blood cell population with Lympholyte®-Rat 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 0.2 μg of this antibody.
  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 (Streptavidin-FITC) 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).
  Results:
  Tissue Distribution by Flow Cytometry Analysis:
  Cell Concentration: 1x10e6 cells per test
  Antibody Concentration Used: 0.2μg/10e6 cells
  Isotypic Control: Biotin Mouse IgG1
  Cell Source:
  Bone Marrow 98.9%
  Blood 47.3%
  Spleen 96.7%