Cd4 Rat Monoclonal Antibody [Clone ID: CT-CD4]

CAT#: CL004B

Cd4 rat monoclonal antibody, clone CT-CD4, Biotin

Product Images

Cell Source: Spleen Percentage of cells stained above control: 18.38%

Specifications

Product Data

• Clone Name: CT-CD4
• Applications: FC
• Recommended Dilution: Flow Cytometry (See Protocols).
  (Reported to be useful in immunohistochemistry on Acetone Fixed Frozen Sections).
• Reactivities: Mouse
• Host: Rat
• Isotype: IgG2a
• Clonality: Monoclonal
• Specificity: This CT-CD4 monolconal antibody (mAb) recognizes Mouse CD4 (L3T4).
• Formulation: PBS containing 0.09% Sodium Azide and EIA grade BSA as a stabilizing protein to bring total protein concentration to 4-5 mg/ml
  Label: Biotin
  State: Liquid purified IgG fraction
• Concentration: lot specific
• Conjugation: Biotin
• Storage: Store the antibody 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.
• Database Link:
  Entrez Gene 12504 Mouse
  UniProt ID P06332
• Background: CD4 (L3T4) which is expressed on the majority of thymocytes and on the MHC class II restricted subset of mature T cells including Th cells1,2. Mouse CD4 has also been reported to be present on multipotential hematopoietic stem cells, bone marrow myeloid precursors, and intrathymic precursors2,3. As a coreceptor in the TCR complex, CD4 is involved in T cell activation through interaction with MHC class II on APC’s and in signal transduction via protein tyrosine kinase lck1.
• Synonyms: T-cell surface antigen T4/Leu-3
• 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 2x10⁷ cells/ml in media A. Add 50 µl of this suspension to each tube (each tube will then contain 1 x 10⁶ cells, representing 1 test).
  4. To each tube, add ~1.0-0.5 µg of this Ab per 10⁶ 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 (Streptavidin-PE) at a 1:20 dilution.
  9. Incubate tubes at 4°C for 30 - 60 minutes (It is recommended that tubes are protected from light since most fluorochromes are light sensitive).
  10. Wash 2 times at 4°C.
  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).
  
  Tissue Distribution by Flow Cytometry Analysis:
  Mouse Strain: BALB/c
  Cell Concentration: 1x10⁶ cells per test
  Antibody Concentration Used: 0.5 µg/10⁶ cells
  Isotypic Control: Biotin Rat IgG2a