CD45/CD14 antibody

Catalog No. ABIN1112151

Product Details

Target: CD45/CD14

Reactivity: Human

Host: Mouse

Clonality: Monoclonal

Conjugate: Un-conjugated

Application: Flow Cytometry (FACS), Immunofluorescence (IF), Immunohistochemistry (IHC)

Characteristics: Monoclonal Mouse anti-human CD45, CD14. The CD45 monoclonal antibody is directed against the CD45- antigen, defined T200 or Leucocyte Common Antigen. The antibody reacts with all cells of the haemopoietic lineage, not with cells of other lineages. The CD14 monoclonal antibody is directed against the CD14- antigen, which is expressed on human monocytes and macrophages. The antibody reacts with human monocytes and macrophages; weak reactions may occur with neutrophils.

Clone: D3-9-47-3D6

Isotype: IgG1

Application Details

Application Notes: It is recommended for use in flow cytometry. This reagent is effective for direct immunofluorescence staining of human tissue for flow cytometric analysis using 20 µl/10^6 cells. Clinical applications: Abnormal numbers of cells expressing the target antigen or cells showing aberrant expression levels of the antigen can be expected in some disease states. This may result in an altered pattern of stainingIt is important to understand the normal expression pattern of an antigen and its relationship to the expression of other relevant antigens in order to perform appropriate analysis. Nonlymphocyte components of LWB that may contaminate the lymphocyte analysis gate include monocytes, granulocytes (neutrophils, eosinophils, basophils), and debris (residual erythrocytes, erythrocyte ghosts, and platelets). The lymphocyte analysis gate should be set to include the largest possible number of lymphocytes in the sample. This is done to avoid biasing the data by selectively excluding lymphocyte subsets exhibiting slightly different lightscatter patterns than the majority of lymphocytes.

Comment:

Fluorescein isothiocyanate (Molecular Probes), R-Phycoerythrin.

Sample Collection: 1. Transfer 100 µl of anticoagulated (EDTA) blood to a 12 x 75 mm polystyrene test tube (10^6 cells). 2. Add 20 µl of CD45 FITC/CD14PE and mix gently with a vortex mixer. The 20 µl is a guideline only, the optimal volume should be determined by the individual laboratory. 3. The recommended negative control is a non-reactive FITC-conjugated antibody of the same isotype. , a non-reactive PE-conjugated antibody of the same isotype 4. Incubate in the dark at room temperature at 4°C for 15 minutes or at room temperature (20-25 °C) for 15 minutes. 5. Add 1,5 ml of Lysing Solution to each sample and mix gently with a vortex mixer. Incubate for 10 minutes at room temperature in the dark. 6. Centrifuge at 1000 x g for 5 minutes. Gently aspirate the supernatant and discard it leaving approximately 50 µl of fluid. 7. Add 2 ml 0.01 mol/l PBS (It betters that it containing 2% bovine serum albumin) and resuspend the cells by using a vortex mixer. 8. Centrifuge at 1000 x g for 5 minutes. Gently aspirate the supernatant and discard it leaving approximately 50 µl of fluid. 9. Resuspend pellet in an appropriate fluid for flow cytometry, e.g. 0.5 ml PBS. The PBS should contain 1% paraformaldehyde (fixative) if samples are not analysed the same day. 10. Analyse on a flow cytometer or store at 2-8 °C in the dark until analysis.

Restrictions: For Research Use only

Handling

Format: Liquid

Buffer: The conjugate is provided in liquid form in buffer containing 1% bovine serum albumin (BSA) and 0,09% Sodium azide, pH 7.2.

Preservative: Sodium azide

Precaution of Use: 1. The device is not intended for clinical use including diagnosis, prognosis, and monitoring of a disease state, and it must not be used in conjunction with patient records or treatment. 2. This product contains Sodium azide (NaN3), a chemical highly toxic in pure form. At product concentrations, though not classified as hazardous, Sodium azide may react with lead and copper plumbing to form highly explosive build-ups of metal azides. Upon disposal, flush with large volumes of water to prevent metal azide build-up in plumbing. 3. As with any product derived from biological sources, proper handling procedures should be used.

Storage: 4 °C

Target Details

Target: CD45/CD14

Background: The CD45 antibody recognizes 180, 195, 205, 220, kDa MW components of the leucocyte common antigen complex to be found on lymphocytes, monocytes, granulocytes, thymocytes and malignant T and B cells. No reactivity has been observed with primary or metastatic carcinoma cells. Plasma cells or myeloma cells may have weak expression or be negative for this antigen. The CD14 antibody recognizes 55kDa MW monocyte surface antigen identified by monoclonal antibodies belonging to the CD14 cluster and found on peripheral blood monocytes and weakly on granulocytes. Monocytic cells, interfollicular dendritic cells and dendritic reticulum cells are stained in sections of lymphoid tissue sections.